Ethical Considerations on Clinical Transformation of MSC-Exo in Tumor Therapy / 中国医学伦理学

In recent years, basic scientific research and clinical studies on mesenchymal stem cell-derived exosome (MSC-Exo) have developed rapidly. MSC-Exo has shown good anti-tumor effects in preclinical and clinical studies. However, due to its diverse sources and functions, MSC-Exo therapy still faces many ethical issues in clinical conversion and application, including the unclear tumor types and mechanisms of action applicable to MSC-Exo, the risk/benefit issues in the process of clinical research, the lack of safety/effectiveness evaluation standards, lax access standards, and the incomplete regulatory systems. The technical guidelines and regulatory policies related to the clinical transformation and application of stem cell therapy are constantly improving. Therefore, to effectively avoid ethical risks, regulate the research and treatment related to the clinical transformation and application of MSC-Exo therapy, and improve the safety and efficacy of MSC-Exo therapy in tumor therapy, it is proposed to deepen clinical research on the relationship between MSC-Exo and tumor regulation, strengthen the risk/benefit analysis, supervise and improve the professional quality of researcher, deeply root in the principle of subjects not to be harmed, construct and perfect the relevant regulatory system, and improve the reviewing ability of the ethics committee, so as to promote the rapid clinical transformation of MSC-Exo and bring good news to cancer patients.

Human umbilical cord mesenchymal stem cell-derived extracellular vesicles enhance the regenerative capability of fibrotic liver / 器官移植

Objective To investigate the role of human umbilical cord mesenchymal stem cell-derived extracellular vesicle (hUC-MSC-EV) in the regeneration of fibrotic liver. Methods C57BL/6 mice were randomly divided into the 70% normal liver resection group (Oil+PHx group), 70% liver fibrosis resection group (CCl4+PHx group) and 70% liver fibrosis resection+mesenchymal stem cell-derived extracellular vesicle (MSC-EV) treatment group (CCl4+PHx+MSC-EV group), with 8 mice in each group. LX-2 cell lines were assigned into the phosphate buffer solution (PBS) group, transforming growth factor (TGF)-β group and TGF-β+MSC-EV group. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) in mice after partial liver resection were detected in each group. The expression levels of liver fibrosis and proliferation-related parameters were analyzed in each group. The messenger RNA (mRNA) expression levels of epidermal growth factor (EGF), fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) in LX-2 cells were detected in each group, and their effects on HGF expression in mouse liver were observed. Results Compared with the Oil+PHx group, the serum levels of AST, ALT and LDH were up-regulated, and the degree of fibrosis was more severe, the positive area of Sirius red and α-smooth muscle actin (α-SMA) staining was larger, and the expression level of α-SMA protein was up-regulated in the CCl4+PHx group. Compared with the CCl4+PHx group, the serum levels of AST, ALT and LDH were decreased, the degree of fibrosis was slighter, the positive area of Sirius red and α-SMA staining was decreased, and the expression level of α-SMA protein was down-regulated in the CCl4+PHx+MSC-EV group, and the differences were statistically significant (all P < 0.05). Compared with the Oil+PHx group, the protein expression levels of Ki67 and proliferating cell nuclear antigen (PCNA) were lower in the CCl4+PHx group. Compared with the CCl4+PHx group, the protein expression levels of Ki67 and PCNA were increased in the CCl4+PHx+MSC-EV group, and the differences were statistically significant (all P < 0.05). Compared with the PBS group, the expression level of CollagenⅠ mRNA in LX-2 cells was increased, the expression level of α-SMA protein was up-regulated and the expression level of HGF protein was decreased in the TGF-β group. Compared with the TGF-β group, the expression level of CollagenⅠ mRNA in LX-2 cells was decreased, the expression levels of HGF mRNA and protein were increased, and the expression level of α-SMA protein was decreased in the TGF-β+MSC-EV group, the differences were statistically significant (all P < 0.05). The expression level of HGF protein in the CCl4+PHx group was lower than that in the Oil+PHx group, whereas the difference was not statistically significant (P > 0.05). The expression level of HGF protein in the CCl4+PHx+MSC-EV group was higher than that in the CCl4+PHx group, and the difference was statistically significant (P < 0.05). Conclusions The regenerative capacity of fibrotic liver is weaker than that of normal liver. hUC-MSC-EV may alleviate liver fibrosis and improve liver regeneration by promoting HGF secretion from actived hepatic stellate cells and effectively enhancing the regenerative capacity of fibrotic liver.

Percutaneous application of allogeneic adipose-derived mesenchymal stem cell in dogs submitted to minimally invasive plate osteosynthesis of the tibia

ABSTRACT Purpose To evaluate clinical outcome following minimally invasive plate osteosynthesis (MIPO) associated with percutaneous transplantation of allogeneic adipose-derived mesenchymal stem cells (AD-MSC) at the tibial fracture site in dogs. Methods Thirty-six dogs presenting with nonarticular complete tibial fracture were included in this study. All fractures were treated by the same MIPO technique. The animals were divided in group 1 (n = 20) received a percutaneous application of 3 × 106 AD-MSC at the fracture site and group 2 (n = 16) did not receive any adjuvant treatment. Postoperative radiographic examinations were made at 15, 30, 60, 90 and 120 days. Results Fifty-eight percent of the patients were classified as skeletally immature. The median weight of the animals was 18.8 kg. The mean radiographic union time differed statistically between the AD-MSC group (28.5 days) and the control group (70.3 days). Sixty percent of dogs in group 1 and 56.25% of the group 2 were considered immature. Conclusions The use of allogeneic AD-MSC cell therapy and MIPO is a safe, viable and effective technique for promoting bone healing in nonarticular tibial fractures in dogs.

Research progress on mesenchymal stem cell in treatment of ischemic-type biliary lesion / 器官移植

At present, surgical and endoscopic interventions are mainly employed to treat ischemic-type biliary lesion (ITBL). Due to the disadvantages of single therapeutic strategy, high difficulty and expensive medical cost, it is urgent to identify a novel treatment option. Mesenchymal stem cell (MSC) has become potential seed cell for tissue and organ repair in regenerative medicine due to its high self-renewal capability, multi-directional differentiation potential, low immunogenicity and immunoregulatory effects, etc. Recent studies have demonstrated that MSC transplantation into ITBL animal models may not only home to the injured area, but also promote the repair of injured biliary tract tissues through anti-apoptotic and pro-angiogenic effect, which indicates that MSC transplantation is expected to become a new strategy for the treatment of ITBL. In this article, the biological characteristics of MSC, the mechanism and clinical application of MSC transplantation for ITBL were reviewed.

Preliminary study of early diagnosis by contrast-enhanced ultrasound combined with mesenchymal stem cell therapy in improving prognosis of biliary ischemia after liver transplantation / 器官移植

Objective To evaluate the clinical efficacy of early diagnosis by contrast-enhanced ultrasound (CEUS) combined with mesenchymal stem cell (MSC) therapy in the treatment of biliary ischemia after liver transplantation. Methods Clinical data of 9 recipients presenting with biliary ischemia detected by CEUS within 4 weeks after liver transplantation and diagnosed with non-anastomotic biliary stricture (NAS) within postoperative 1 year were retrospectively analyzed. In the conventional treatment group, 4 recipients were treated with conventional treatment including liver protection, cholagogic therapy and interventional therapy. In MSC treatment group, 5 recipients received intravenous infusion of MSC at 1, 2, 4, 8, 12 and 16 weeks after biliary ischemia detected by CEUS on the basis of conventional therapy. The interventional treatment and clinical prognosis within 1 year after liver transplantation were analyzed between two groups. Results Two recipients in the MSC treatment group required interventional therapy, which was initially given at 7-9 months after liver transplantation for 1-2 times. All recipients in the conventional treatment group required interventional therapy, which was initially delivered at postoperative 1-3 months for 2-6 times, earlier than that in the MSC treatment group. Within 1 year following liver transplantation, diffuse bile duct injury occurred in 2 recipients in MSC treatment group, and no graft dysfunction was observed. In the conventional treatment group, all recipients developed diffuse bile duct injury, and 2 recipients presented with graft dysfunction. Conclusions Early diagnosis of biliary ischemia after liver transplantation by CEUS combined with MSC therapy may delay and reduce the requirement of interventional therapy for NAS, and also improve clinical prognosis of the recipients.

Mechanism of human umbilical cord mesenchymal stem cells alleviating ischemia-reperfusion injury of hepatocytes through mitochondrial transfer / 器官移植

Objective To explore the mechanism of human umbilical cord mesenchymal stem cell (HUC-MSC) alleviating ischemia-reperfusion injury (IRI) of liver cells through mitochondrial transfer. Methods Normal human liver cell line L02 was divided into the blank control group, oxygen-glucose deprivation (OGD) group, experimental control group, and L02 and HUC-MSC co-culture group (L02+HUC-MSC group). L02+HUC-MSC group was further divided into 10:1 co-culture subgroup (group A), 4:1 co-culture subgroup (group B), 2:1 co-culture subgroup (group C), 1:1co-culture subgroup (group D) and 1:2 co-culture subgroup (group E) according to different co-culture ratio of L02 and HUC-MSC. The apoptosis rate and relative reactive oxygen species (ROS) level of L02 cells were detected by flow cytometry. The MitoTracker positive rate of L02 cells was detected by flow cytometry. The mitochondrial transfer from HUC-MSC to L02 cells was observed by laser confocal microscope. Results The apoptosis rate and relative ROS level of L02 cells in the OGD group were significantly higher than those in the blank control group (both P < 0.05). Compared with the OGD group, the apoptosis rates of L02 cells in group B, C, D and E were significantly decreased (all P < 0.05), and the relative ROS level of L02 cells in group E was significantly declined (P < 0.05). The MitoTracker positive rate of L02 cells did not significantly differ between group A and experimental control group (P>0.05), whereas the MitoTracker positive rates of L02 cells in group B, C, D and E were significantly higher than that in the experimental control group in a concentration-dependent manner (all P < 0.05). Under the laser confocal microscope, mitochondrial transfer fromHUC-MSC to L02 cells could be observed through tunneling nanotube (TNT). Conclusions HUC-MSC may alleviate cell apoptosis and reduce ROS level of liver cells after IRI via direct mitochondrial transfer between cells.

Reduction of recurrent tendinitis scar using autologous mesenchymal stem cells derived from adipose tissue from the base of the tail in Holsteiner horses (Equus ferus caballus) / Reducción de la cicatriz de tendinitis recidivante mediante células madre mesenquimales autólogas derivadas de tejido adiposo de la base de la cola en equinos Holsteiner (Equus ferus caballus)

As a result of their intense physical activity, racehorses suffer high tendon stress which may result in various pathologies. One of these is tendonitis in the tendon of the superficial digital flexor muscle (TSDFM). Conventional treatment with rest, has not shown to be very effective, and regenerative medicine through the application mesenchymal stem cells appears to be a promising therapy. The objective of this work was to assess the effect of the application of autologous MSC on reduction of the scar length in recurrent TSDFM tendinitis in Holsteiner horses, using image analysis. This study included two groups of five animals each: A control group that received conventional treatment (CG) and an experimental group which was also treated with intralesional injections of MSC (EG). Scar evolution was assessed by echographic analysis, with measurements taken of the scar length over a four month period; the length at month zero, was taken as the initial value of 100 %. During the first month, the mean scar length diminished to 81.14 % (EG) and 95.85 % (CG); after the second month, lengths were 64.4 % (EG) and 92.3 % (CG); following the third month lengths were 51.92 % (EG) and 87.42 % (CG); finally at the end of the fourth month the lengths recorded were 26.7 % (EG) and 83.92 % (CG). These results show that treatment with autologous MSC helps TSDFM scar length was significantly reduced, as compared to conventional treatment.

Reducción de la cicatriz de tendinitis recidivante mediante células Madre mesenquimales autólogas derivadas de tejido adiposo de la base de la cola en equinos Holsteiner (Equus ferus caballus). En equinos deportistas, la actividad física intensa ocasiona gran estrés en los tendones, pudiendo ocasionar diversas patologías como la tendinitis del tendón del músculo flexor digital superficial (TMFDS). El tratamiento convencional con reposo es poco eficaz, siendo la medicina regenerativa a través de la aplicación de células madres mesenquimáticas (MSC) una promisoria terapia. El objetivo de este trabajo, fue evaluar el efecto de la aplicación de MSC autólogas, sobre la reducción de la longitud de la cicatriz en tendinopatías recidivantes del TMFDS en equinos Holsteiner, a través del análisis de imagen. Este estudio conto con dos grupos de cinco animales cada uno, el grupo control mantuvo el tratamiento convencional (GC) y el grupo experimental fue tratado adicionalmente con inyección interlesional de MSC (GE). El análisis ecográfico permitió evaluar la evolución de la cicatriz, a través de la medición de su longitud durante los cuatros meses, tomando la longitud del mes cero como la medición inicial del 100 %. Durante el primer mes, la longitud de la cicatriz se redujo a un 81,14 % (GE) y 95,85 % (GC), al segundo mes la longitud fue de un 64,4 % (GE) y de 92,3 % (GC), al tercer mes, la longitud fue de 51,92 (GE) y un 87,42 (GC), finalmente al cuarto mes la longitud fue de 26,7 % (GE) y del 83,92 % (GC). Estos resultados muestran que el tratamiento con MSC autólogas favorece a la disminución de la longitud de la cicatriz del TMFDS de forma significativa respecto al tratamiento convencional.

Mesenchymal Stem Cell Transplantation Promotes Functional Recovery through MMP2/STAT3 Related Astrogliosis after Spinal Cord Injury

BACKGROUND AND OBJECTIVES: Treatment with mesenchymal stem cells (MSC) in spinal cord injury (SCI) has been highlighted as therapeutic candidate for SCI. Although astrogliosis is a major phenomenon after SCI, the role of astrogliosis is still controversial. In this study, we determined whether acute transplantation of MSC improves the outcome of SCI through modulating astrogliosis. METHODS: Bone marrow derived rat MSCs were induced neural differentiation and transplanted after acute SCI rats. Matrix metalloproteinase (MMP) and neuro-inflammatory pathway were analyzed for acute astrogliosis at 1, 3 and 7 d after SCI in RT-PCR- and western blot analysis. Functional outcome was assessed serially at postoperative 1 d and weekly for 4 weeks. Histopathologic analysis was undertaken at 7 and 28 d following injury in immunohistochemistry. RESULTS: Transplantation of MSCs decreased IL-1α, CXCL-2, CXCL-10, TNF-α and TGF-β in a rat model of contusive SCI. Protein level of NF-κB p65 was slightly decreased while level of STAT-3 was increased. In immunohistochemistry, MSC transplantation increased acute astrogliosis whereas attenuated scar formation with increased sparing white matter of spinal cord lesions. In RT-PCR analysis, mRNA levels of MMP2 was significantly increased in MSC transplanted rats. In BBB locomotor scale, the rats of MSC treated group exhibited improvement of functional recovery. CONCLUSIONS: Transplantation of MSC reduces the inflammatory reaction and modulates astrogliosis via MMP2/STAT3 pathway leading to improve functional recovery after SCI in rats.

CRISPR/Cas9 Edited sRAGE-MSCs Protect Neuronal Death in Parkinson's Disease Model

BACKGROUND AND OBJECTIVES: Parkinson’s disease (PD) is a fatal and progressive degenerative disease of the nervous system. Until recently, its promising treatment and underlying mechanisms for neuronal death are poorly understood. This study was investigated to identify the molecular mechanism of neuronal death in the substantia nigra and corpus striatum of PD. METHODS: The soluble RAGE (sRAGE) secreting Umbilical Cord Blood—derived Mesenchymal Stem Cell (UCB-MSC) was generated by gene editing method using clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9 (CRISPR/Cas9). These cells were transplanted into Corpus Striatum of rotenone-induced PD animal models then behavioral test, morphological analysis, and immunohistochemical experiments were performed to determine the neuronal cell death and recovery of movement. RESULTS: The neuronal cell death in Corpus Striatum and Substantia Nigra was dramatically reduced and the movement was improved after sRAGE secreting UCB-MSC treatment in PD mice by inhibition of RAGE in neuronal cells. CONCLUSIONS: We suggest that sRAGE secreting UCB-MSC based therapeutic approach could be a potential treatment strategy for neurodegenerative disease including PD.

Stable Knockout of lncRNA DANCR Gene in Mesenchymal Stem Cells with a Dual-Target CRISPR Lentiviral Vector / 中山大学学报(医学科学版)

@#【Objective】Using the CRISPR/Cas9（CRISPR/crispr-associated（Cas）9 method，a dual-target lentiviral vector containing single- guide RNAs（sgRNAs）targeting both the 5’and 3’ends of the anti- differentiation noncoding RNA（DANCR）gene was constructed. Stable knockout of DANCR gene in mesenchymal stem cells（MSC）would be helpful for the future study of the biological function of DANCR.【Methods】Designed sgRNAs targeting either the 5’or 3’ end of DANCR and cloned into two CRISPR vectors. The vector was transfected into 293FT cells，and the genomic DNA of 293FT cells was extracted to verify the efficiency of individual sequence. Two functional sgRNAs targeting either the 5’ or 3’end were incorporated into a same lentiviral CRISPR vector through gateway and enzymatic ligation. 293FT was used for lentiviral packaging，after which the virus was harvested to infect MSC，and the knockout efficiency of DANCR in MSC was detected.【Results】All four sgRNA sequences targeting DANCR successfully guided Cas9 to cleave the gene. sgRNAs targeting either the 5’and 3’end were combined to establish a dual-target lentiviral vector for stable knockout of DANCR. The vector was packaged into lentivirus and infected MSC. Finally，we successfully obtained mesenchymal stem cell lines with DANCR gene knockout.【Conclusions】Using the CRISPR method，a dual-target lentiviral vector can efficiently and stably knock out DANCR gene in MSC.

Catharmus tinctorius volatile oil promote the migration of mesenchymal stem cells via ROCK2/Myosin light chain signaling / 中国天然药物

MSC transplantation has been explored as a new clinical approach to stem cell-based therapies for bone diseases in regenerative medicine due to their osteogenic capability. However, only a small population of implanted MSC could successfully reach the injured areas. Therefore, enhancing MSC migration could be a beneficial strategy to improve the therapeutic potential of cell transplantation. Catharmus tinctorius volatile oil (CTVO) was found to facilitate MSC migration. Further exploration of the underlying molecular mechanism participating in the pro-migratory ability may provide a novel strategy to improve MSC transplantation efficacy. This study indicated that CTVO promotes MSC migration through enhancing ROCK2 mRNA and protein expressions. MSC migration induced by CTVO was blunted by ROCK2 inhibitor, which also decreased myosin light chain (MLC) phosphorylation. Meanwhile, the siRNA for ROCK2 inhibited the effect of CTVO on MSC migration ability and attenuated MLC phosphorylation, suggesting that CTVO may promote BMSC migration via the ROCK2/MLC signaling. Taken together, this study indicates that C. tinctorius volatile oil could enhance MSC migration via ROCK2/MLC signaling in vitro. C. tinctorius volatile oil-targeted therapy could be a beneficial strategy to improve the therapeutic potential of cell transplantation for bone diseases in regenerative medicine.

Reorganization of Actin Cytoskeleton and Microtubule Array during the Chondrogenesis of Bovine MSCs

Mesenchymal stem cells (MSCs) are multipotent stem cells that are capable of self-renewal and can be committed into classical mesodermal tri-lineage differentiation (adipocytes, osteocytes and chondrocytes). During chondrogenic differentiation MSCs change their shape due to the reorganization of cytoskeletal components. This has been well documented for human and rodent models. Morphological changes of microtubule network and actin filaments that occur during the chondrogenic differentiation of MSCs from large animal models remain unknown. In this study we described the morphological changes of cell shape, area, actin structures and microtubule array that occur in bovine MSCs during the chondrogenic differentiation of bovine bone-marrow isolated MSCs. Chondrogenic differentiation of bMSCs occur more rapidly on glass substrate compared to the cells plated on vitronectin, and in 7 days after the commitment we observed clusters of small round-shaped cells that expressed glycosaminoglycans. During the differentiation microtubule (MT) array of MSCs became non-radial, and non-centrosomal MTs that grew transversely to the cell radius appeared in the inner cytoplasm and near the cell edges. At the end of differentiation process we observed the thick bundles of MTs that grew in parallel to the cell edge and basket-like structures of curved MTs around the nucleus. The main changes of actin structures in differentiating MSCs included the disappearance of thick transverse stress fibers and actin arches and reorganization of actin into chaotic network of thin cortical fibers. Our results imply the important role of both actin and MT cytoskeletal systems in chondrogenesis and reveals new perspectives for experimental regulation of these process in vitro systems.

Nuevas áreas de vida ­ Revisión muerte súbita 2018 / New areas of life ­ Sudden death review 2018

Una de cada cinco muertes en adultos en países desarrollados se debe a causas cardiovasculares; la mitad de esas muertes se produce de forma súbita y un gran porcentaje en el ámbito extrahospitalario. Las medidas de prevención se dividen en: aquellas destinadas a prevenir en primer lugar que el evento de muerte súbita cardíaca suceda, y aquellas cuyo objetivo es actuar en el momento en que el evento de muerte súbita está sucediendo. Las primeras tienen como objetivo disminuir las principales causas de muerte súbita en países desarrollados: las cardiopatías estructurales (cuya principal causa es la enfermedad coronaria). En este sentido, con el fin de intentar paliar el desarrollo de una cardiopatía que predisponga a la aparición de arritmias fatales y la MSC, se implementan medidas de prevención primarias higiénico-dietéticas y farmacológicas (con el objetivo de disminuir y el controlar los factores de riesgo) y, en aquellos con enfermedad cardiovascular ya establecida, se implementan las estrategias secundarias farmacológicas y/o quirúrgicas (revascularización, reemplazo valvular, etc.). El segundo abordaje surge del hecho de que, a pesar de todas estas medidas, un gran número de pacientes presentará eventos arrítmicos en el ámbito extrahospitalario (MSCEH), ya sea porque aunque recibieron el tratamiento óptimo presentan aún un elevado riesgo de MSC, porque no fueron diagnosticados a tiempo o porque a pesar de haber hecho estudios complementarios el diagnóstico es muy dificultoso. Existen dos estrategias: la primera son los dispositivos de cardiodesfibrilación implantables (o, más recientes, los chalecos vestibles). Estos aparatos están indicados para una población seleccionada, sea por haber presentado ya un episodio de muerte súbita abortado, o por presentar una cardiopatía (estructural o genética) que predisponga a una mayor probabilidad de sufrir un evento. La segunda estrategia es la educación y el desarrollo de programas de salud pública que permitan capacitar a la población general en la realización de RCP y el uso de desfibriladores automáticos externos (DEAs), los cuales deberían estar disponibles en cualquier lugar público. Múltiples estudios demostraron que el acceso de la población general al aprendizaje de maniobras de RCP sencillas y pragmáticas y la presencia de DEAs se traduce en un gran aumento de sobrevida sin secuelas en víctimas de MSCEH. (AU)

One of every five deaths in adults is due to cardiovascular causes, in developed countries, and half of these deaths will occur suddenly. A large percentage occur in the out of hospital setting, so measures to prevent it are divided into: those designed to prevent, in the first place, the sudden cardiac death event from happening and those whose purpose is to act when the sudden death event that has already occurred and it´s ongoing. The first aims to reduce the main causes of sudden death in developed countries: structural heart disease (with coronary heart disease as its main cause). In this regard, with the purpose to mitigate the development of a heart disease that predisposes the occurrence of fatal arrhythmias and SCD, we have primary prevention measures, like healthy life style conduct with or without pharmacological treatment, (whose objective is the reduction and control of cardiovascular risk factors) and, in those with cardiovascular disease already established, there is an implementation of pharmacological and / or surgical strategies (Revascularization, valve replacement, etc.). The second objective arises from the fact that, despite all these preventive and therapeutic measures, a large number of patients will present out-of-hospital cardiac arrest (OHCA) either because although they received optimal treatment they still remain in high risk of SCD, even because they were not diagnosed on time, or because despite having complementary studies made the diagnosis is very difficult. There are two well strategies: the first are implantable cardio-defibrillation devices (or, more recently, wearable vests). These are indicated for a selected population, either because they have already presented an episode of sudden aborted death, or because they have heart disease (structural or genetic), which predisposes to a greater probability of suffering an event. The second strategy is the education and development of public health programs that enable the general population to be trained in CPR and the use of external automatic defibrillators. (AEDs) should be available in any public place. Multiple studies showed that access to the general population for learning simple and pragmatic CPR maneuvers and the presence of AEDs is making an impact on a significant increase in survival without consequences in OHCA victims. (AU)

Análises macroscópica e histopatológica do alotransplante parcial de vesícula urinária com células-tronco mesenquimais alogênicas derivadas do tecido adiposo em coelhos / Macroscopic and histopathological analysis of partial urinary bladder allograft using allogenic adipose tissue derived mesenchymal stem cells in rabbits

O número de transplantes de órgãos e tecidos em humanos e animais tem crescido significativamente nos últimos anos, principalmente após o advento de técnicas modernas e mais seguras indutoras de imunossupressão. Objetiva-se com o presente estudo avaliar macro e microscopicamente o alotransplante parcial de bexiga a fresco em coelhos, utilizando como agente imunomodulador células-tronco mesenquimais derivadas do tecido adiposo (ADSC) alogênicas. Foram utilizados 25 coelhos, sendo um deles macho e doador das ADSCs, e os outros 24 eram fêmeas, submetidas a alotransplante parcial de bexiga, tratadas com ciclosporina (GCi) ou células-tronco mesenquimais (GCe). Conclui-se que o GCe teve melhor aceitação histológica do implante em relação ao GCi aos 30 dias de avaliação.(AU)

The number of organ and tissue transplantation in humans and animals has grown significantly recently, especially after the advent of modern and safer techniques of immunosuppression. The objective of this study was to evaluate macro and microscopically partial urinary bladder fresh allograft in rabbits, using as immunomodulatory agent cyclosporine or allogenic adipose tissue derived mesenchymal stem cells (ADSCs). For this purpose, 25 rabbits were used. One male was the donor of ADSCs; 24 females received a partial urinary bladder allograft and were treated with cyclosporine (GCi) or mesenchymal stem cells (GCe). We conclude that the GCe group had better histological acceptance of the implant than GCi group at 30 days evaluation.(AU)

Characteristics and clinical significance of osteogenic differentiation of mesenchymal stem cells derived from bone marrow in patients with myelodysplastic syndromes / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the osteogenic differentiation characteristics of mesenchymal stem cell (MSC) derived from bone marrow in patients with myelodysplastic syndromes (MDS) and its clinical significance. Methods: Bone marrow samples from 30 cases of newly diagnosed untreated MDS patient atAffiliated Hospital of Heibei University were collected for this study. MSCs from MDS patients and normal subjects were isolated and cultured, and morphological characteristics of MSCs were observed in vitro; under proper conditions, MSCs were induced to differentiate into osteoblasts and adipocytes; The formation of calcium nodules at 14th day after osteogenic differentiation was observed by alizarin red staining; mRNA expressions of osteogenic differentiation transcription factors Ostefix and RUNX2 in undifferentiated MSCs, as well as the mRNAexpression of Jagged-1, which involved in the transformation from hematopoietic cells into leukemic cells, were detected by quantitative PCR. Results: The MSCs derived from patients with MDS were characterized with increased cell volume and decreased differentiation potential. Compared with the control group, the expression levels of osteogenic differentiation transcription factors Osterix and RUNX2 were significantly decreased (P < 0.05). Alizarin red staining showed that the content of calcium nodules in MDS group was significantly less than that in the normal control group, while the expression level of Jagged-1 was significantly higher (P < 0.05). Conclusion: MSCs derived from bone marrow of MDS patients showed significant increased cell volume, decreased differentiation potential and elevated Jagged-1 expression; all of these might play important roles in the .hematopoietic failure and progression to acute myeloid leukemia in MDS patients.

Effects of SphK1 gene on the proliferation and migration of colon cancer RKO cells induced by mesenchymal stem cells / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of sphingosine kinase 1 (SphK1) knockdown on the proliferation and migration of colon cancer RKO cells induced by mesenchymal stem cells (MSCs). Methods: RKO cells were treated with MSCs conditioned medium (MSC-CM) or control medium (Control-CM), respectively. Cell proliferation was detected by CCK-8 assay. Cell migration ability was tested by Transwell chamber assay. The proteins expression of Ki-67, MMP-2/9, CD44 and CD133 was detected by Western blotting. Then, the expression of SphK1 in RKO cells was suppressed by targeted gene lentivirus shRNA vector transfection. The effects of SphK1 knockdown on the proliferation, migration and protein expressions of Ki-67, MMP-2/9, CD44 and CD133 of RKO cells induced by MSC-CM were observed. Results: The RKO cells proliferation was promoted by MSC-CM in a time-dependent manner; moreover (P<0.05), the migration ability of cells was significantly enhanced after being treated with MSC-CM(P<0.01). In addition, MSC-CM significantly increased the protein expressions of Ki-67, MMP-2/9, CD44 and CD133(all P<0.05 or P<0.01). Lentiviral ShRNA vector transfection could significantly inhibit the expression of SphK1. Down-regulation of SphK1 significantly inhibited the proliferation, migration and protein expressions of Ki-67, MMP-2/9, CD44 and CD133 of RKO cells induced by MSC-CM(all P<0.05 or P<0.01). Conclusion: MSC-CM promotes the proliferation and migration of colon cancer RKO cells. Down-regulation of SphK1 reverses the cell proliferation and migration induced by MSC-CM via inhibiting the expression of MMP-2/9, CD44 and CD133.

Isolation, culture and characterization of multipotent mesenchymal stem cells from goat umbilical cord blood / Isolamento, cultura e caracterização de células tronco mesenquimais multipotentes provenientes do sangue do cordão umbilical caprino

Mesenchymal stem cells (MSC) reside in small numbers in many adult tissues and organs, and play an active role in the homeostasis of these sites. Goat derived multipotent MSC have been established from bone marrow, adipose tissues and amniotic fluid. Umbilical cord blood (UCB) is considered an important source of these cells. However, the MSC isolation from the goat UCB has not been demonstrated. Therefore, the aim of the present study was to isolate, culture and characterize goat umbilical cord blood derived mesenchymal stem cells. MSC were isolated from UCB by Ficoll-Paque density centrifugation and cultured in DMEM supplemented with 10% or 20% FBS. FACS analysis was performed and induction lineage differentiation was made to characterize these cells. They exhibited two different populations in flow cytometry, and revealed the positive expression of CD90, CD44 and CD105, but negative staining for CD34 in larger cells, and positive stained for CD90 and CD105, but negative for CD44 and CD34 in the smaller cells. MSC from goat UCB showed capability to differentiate into chondrocytes and osteoblasts when incubated with specific differentiation medium. Present study established that goat mesenchymal stem cells can be derived successfully from umbilical cord blood.(AU)

As células tronco mesenquimais (MSC) residem em pequenas quantidades em muitos tecidos e órgãos adultos, desempenhando um papel ativo na homeostase destes locais. O isolamento de MSC já foi demonstrado em amostras de medula óssea, tecido adiposo e fluido amniótico de cabras. O sangue de cordão umbilical é considerado uma fonte importante desse tipo de células. No entanto, até o presente momento, não foi demonstrado o isolamento de MSC provenientes do sangue de cordão umbilical de cabras. Dessa forma, o objetivo do presente estudo foi isolar, cultivar e caracterizar células tronco mesenquimais provenientes do sangue do cordão umbilical caprino. As MSC foram isoladas utilizando o gradiente de densidade Ficoll-Paque e cultivadas em DMEM suplementado com 10% ou 20% de FBS. A caracterização desse tipo celular foi realizada através de análise por citometria de fluxo e diferenciação em linhagens celulares mesodermais. A analise no citômetro de fluxo demonstrou a presença de duas populações distintas, um grupo com células maiores e outro com células menores; observando expressão positiva de CD90, CD44 e CD105, e negativa para CD34 nas células maiores; enquanto que as menores foram positivas para CD90 e CD105, mas negativas para CD44 e CD34. As células isoladas demonstraram capacidade de se diferenciar em condrócitos e osteoblastos quando incubadas com meio de diferenciação específico. O presente estudo demonstrou que células tronco mesenquimais podem ser obtidas com sucesso do sangue do cordão umbilical caprino.(AU)

Comparación entre Células Troncales Mesenquimales obtenidas de Médula Ósea, Tejido Adiposo y Gelatina de Wharton en base a los Criterios de la ISCT / Comparison between Mesenchymal Stem Cells obtained from Bone Marrow, Adipose Tissue and Wharton Gelatin according to the ISCT Criteria

RESUMEN: Las células troncales mesenquimales (CTM) representan una población heterogénea con capacidad para auto-renovarse y diferenciarse a distintos tipos celulares. Estas fueron descritas en un inicio en médula ósea (MO) a mediados del siglo pasado, desde entonces este tejido se ha convertido en el estándar de oro para la obtención y caracterización de CTM. Actualmente se sabe que este tipo de células se encuentran alojadas en nichos distribuidos por todo el organismo, donde contribuyen a los procesos de regeneración del tejido donde se localizan. No obstante, encontrar una fuente alterna de CTM con las mismas características que las de MO, pero que su extracción no suponga riesgo para el donador es fundamental para su utilización con fines terapéuticos. En este trabajo se aislaron células troncales de médula ósea, y se compararon con tejido adiposo y gelatina de Wharton y caracterizaron de acuerdo a los criterios de la Sociedad Internacional para la Terapia Celular (ISCT). Los resultados mostraron que la morfología, diferenciación osteogénica y adipogénica, así como la expresión de los antígenos de superficie CD90, CD73 y CD105 cumplen con los estándares, señalando a las provenientes de gelatina de Wharton como mejor opción.

ABSTRACT: Mesenchymal stem cells (MSC) represent a heterogeneous population with the capacity to self-renew and differentiate into different cell types. At the middle of the last century these cells initially were described in bone marrow (BM), thence this tissue has become the gold standard for obtaining and characterization of MSC. It is known that these cells are housed in specific areas called niches distributed throughout all body, where they contribute to tissue regeneration processes of self-tissue were they are located. However, finding an alternative source of CTM with the same characteristics that have showed in MO, but its obtention no represent a risk since the donor is essential to their use for therapeutic purposes. In this study we isolated mesenchymal stem cells from bone marrow, adipose tissue and Wharton's jelly and they were compared in their characteristics in according to the standards of the International Society for Cellular Therapy (ISCT). The results showed that the morphology as well as adipogenic and osteogenic differentiation and also the expression of surface antigens (CD90, CD73, and CD105) from all tissues accomplished the standards, although Wharton's jelly represented the best option.

Goat umbilical cord cells are permissive to small ruminant lentivirus infection in vitro

Abstract Small ruminant lentiviruses isolated from peripheral blood leukocytes and target organs can be propagated in vitro in fibroblasts derived from goat synovial membrane cells. These cells are obtained from tissues collected from embryos or fetuses and are necessary for the establishment of the fibroblast primary culture. A new alternative type of host cells, derived from goat umbilical cord, was isolated and characterized phenotypically with its main purpose being to obtain cell monolayers that could be used for the diagnosis and isolation of small ruminant lentiviruses in cell culture. To accomplish this goal, cells were isolated from umbilical cords; characterized phenotypically by flow cytometry analysis; differentiate into osteogenic, chondrogenic and adipogenic lineage; and submitted to viral challenge. The proliferation of goat umbilical cord cells was fast and cell monolayers formed after 15 days. These cells exhibited morphology, immunophenotype, growth characteristics, and lineage differentiation potential similar to mesenchymal stem cells of other origins. The goat umbilical cord derived cells stained positive for vimentin and CD90, but negative for cytokeratin, CD34 and CD105 markers. Syncytia and cell lysis were observed in cell monolayers infected by CAEV-Cork and MVV-K1514, showing that the cells are permissive to small ruminant lentivirus infection in vitro. These data demonstrate the proliferative competence of cells derived from goat umbilical cords and provide a sound basis for future research to standardize this cell lineage.

The Therapeutic Efficacy of Tonsil-derived Mesenchymal Stem Cells in Dextran Sulfate Sodium-induced Acute Murine Colitis Model / 대한소화기학회지

BACKGROUND/AIMS: Mesenchymal stem cells (MSCs) are multipotent progenitor cells currently under investigation for its efficacy as the treatment for inflammatory bowel disease. In this study, we evaluated the efficacy of tonsil-derived mesenchymal stem cells (T-MSCs) as a novel source of mesenchymal stem cells and traced their localization in a murine model of acute colitis induced by dextran sulfate sodium (DSS). METHODS: C57BL/6 mice were randomly assigned to the following three groups: the normal control group, DSS colitis group (DSS+phosphate buffered saline), and T-MSC group (DSS+T-MSCs, 1×106). The severity of colitis was assessed by determining the severity of symptoms of colitis, colon length, histopathologic grade, and levels of inflammatory cytokines. T-MSCs labeled with PKH26 were traced in vivo. RESULTS: The T-MSC group, compared with the DSS colitis group, showed a significantly lower disease activity index (11.3±1.5 vs. 8.3±1.9, p=0.015) at sacrifice and less reduction of body weight (-17.1±5.0% vs. -8.1±6.9%, p=0.049). In the T-MSC group, the histologic colitis score was significantly decreased compared with the DSS colitis group (22.6±3.8 vs. 17.0±3.4, p=0.039). IL-6 and IL-1β, the pro-inflammatory cytokines, were also significantly reduced after a treatment with T-MSCs. In vivo tracking revealed no PKH26-labelled T-MSCs in the colonic tissue of mice with acute colitis. CONCLUSIONS: In the acute colitis model, we demonstrated that the administration of T-MSCs ameliorates inflammatory symptoms and histology. Moreover, the anti-inflammatory activities of T-MSCs were independent of gut homing.