Stimulation of mouse hematopoietic stem cells by angiogenin and DNA preparations

Immature hematopoietic progenitors are a constant source for renewal of hemocyte populations and the basic component of the tissue and cell repair apparatus. A unique property of these cells of internalizing extracellular double-stranded DNA has been previously shown. The leukostimulatory effect demonstrated in our pioneering studies was considered to be due to the feature of this cell. In the present research, we have analyzed the effects of DNA genome reconstructor preparation (DNAgr), DNAmix, and human recombinant angiogenin on both hematopoietic stem cells and multipotent progenitors. Treatment with bone marrow cells of experimental mice with these preparations stimulates colony formation by hematopoietic stem cells and proliferation of multipotent descendants. The main lineage responsible for this is the granulocyte-macrophage hematopoietic lineage. Using fluorescent microscopy as well as FACS assay, co-localization of primitive c-Kit- and Sca-1-positive progenitors and the TAMRA-labeled double-stranded DNA has been shown. Human recombinant angiogenin was used as a reference agent. Cells with specific markers were quantified in intact bone marrow and colonies grown in the presence of inducers. Quantitative analysis revealed that a total of 14,000 fragment copies of 500 bp, which is 0.2% of the haploid genome, can be delivered into early progenitors. Extracellular double-stranded DNA fragments stimulated the colony formation in early hematopoietic progenitors from the bone marrow, which assumed their effect on cells in G0. The observed number of Sca1+/c-Kit+ cells in colonies testifies to the possibility of both symmetrical and asymmetrical division of the initial hematopoietic stem cell and its progeny.

Chronic Injury of Mice Bone Marrow Multipotent Hematopoietic Progenitor Cells Induced by Ionizing Radiation / 中国实验血液学杂志

OBJECTIVE@#To explore the chronic injury and its possible mechanism of ionizing radiation on multipotent hematopoietic progenitor cells (MPPs) by determining the related indicators of MPPs in bone marrow of mice post-radiation.@*METHODS@#Sixteen C57BL/6 adult mice were randomly divided into normal control and irradiation groups, 8 mice in each group. The mice in irradiation group were exposed to 6 Gy X-ray. The proportion of bone marrow MPPs, their apoptosis and proliferation 2 months after irradiation were detected by flow cytometry. Mitochondrial activity and levels of reactive oxygen species (ROS) in each MPPs population were detected by Mitotracker Red and DCFDA probes, and the senescent state of MPPs in the bone marrow was analyzed.@*RESULTS@#Ionizing radiation could reduce the proportion of MPPs in mouse bone marrow. The proportions and numbers of MPP1, MPP3 and MPP4 in the bone marrow were significantly decreased after whole-body irradiation with 6 Gy X-ray (P<0.05). In addition, radiation significantly reduced the colony-forming capacity of MPPs in bone marrow (P<0.05), the proportions of apoptotic cells in the MPP1 and MPP4 cell populations increased significantly in the bone marrow (P<0.05). The activity of mitochondria was significantly reduced in the bone marrow MPP2, MPP3 and MPP4 cell populations compared with that of the control group (P<0.05). It was also found that the radiation could significantly increase the ROS levels of MPPs in bone marrow, and the content of ROS in the MPP2, MPP3 and MPP4 cell population of the bone marrow was significantly increased(P<0.05). The senescent cells ratios of MPP1, MPP3 and MPP4 cells in the bone marrow after irradiation were significantly higher than those in the control group (P<0.05).@*CONCLUSION@#Ionizing radiation can cause chronic MPPs damage in mice, which is closely associated with persistent oxidative stress, cells apoptosis, and cellular senescence.

Efficiency and safety of umbilical cord mesenchymal stromal/ stem cells in multiple sclerosis combined treatment / Eficacia y seguridad de las células madre / estromales mesenquimales del cordón umbilical en el tratamiento combinado de la esclerosis múltiple

SUMMARY: Multiple sclerosis is a demyelinating disease of the nervous system that affects young people of working age and quickly leads to disability. Treatment of this pathology with umbilical cord mesenchymal stem cells is promising, given their immunomodulatory and neurotrophic properties. The study involved 27 patients diagnosed with multiple sclerosis, 12 of whom underwent combined treatment (intravenous and intrathecal administration) of umbilical cord multipotent mesenchymal stromal/stem cells. The effectiveness of treatment was determined by the degree of neurological deficit and spasticity. Combined treatment with umbilical cord mesenchymal stem cells significantly improves the condition of patients with multiple sclerosis and promotes the regression of neurological deficits and spasticity. This treatment is safe, but for a deeper study, it is necessary to continue research in this area.

RESUMEN: La esclerosis múltiple es una enfermedad desmielinizante del sistema nervioso que afecta a los jóvenes en edad laboral y conduce rapidamente a la discapacidad. El tratamiento de esta patología con células madre mesenquimales de cordón umbilical es prometedor, debido a sus propiedades inmunomoduladoras y neurotróficas. En el estudio participaron 27 pacientes diagnosticados de esclerosis múltiple, 12 de los cuales fueron sometidos a un tratamiento combinado (administración intravenosa e intratecal) de células madre / estromales mesenquimales multipotentes del cordón umbilical. La efectividad del tratamiento estuvo determinada por el grado de déficit neurológico y espasticidad. El tratamiento combinado con células madre mesenquimales del cordón umbilical mejora significativamente la condición de los pacientes con esclerosis múltiple y promueve la regresión de los déficits neurológicos y la espasticidad. Este tratamiento es seguro, sin embargo, es necesario continuar investigando en esta area.

Effect of macrophages on the differentiation of mouse induced pluripotent stem cells into hepatic progenitor cells / 临床肝胆病杂志

ObjectiveTo investigate the effect of macrophages (MCs) on the differentiation of mouse induced pluripotent stem cells (iPSCs) into hepatic progenitor cells (HPCs). MethodsA total of 24 C57BL/6N mice were used to obtain MCs by peritoneal irrigation, and the supernatant was collected to obtain the conditioned medium of MCs (MC-CDM). Activin A, bone morphogenetic protein 4, and fibroblast growth factor were used to induce the differentiation of mouse iPSCs into HPCs. The differentiation of HPCs were randomly divided into control group (normal medium) and experimental group (MC group; use of MC-CDM medium on day 5 of induction). Morphology, immunofluorescence assay, and Western blot were used to compare the morphology of HPCs and the expression of related proteins between the control group and the MC group. The t-test was used for comparison of continuous data between two groups. ResultsHPCs derived from iPSCs were established in vitro, and HPCs had the potential to differentiate into hepatocytes. Immunofluorescence assay showed that compared with the D12 control group, the D12 MC group had a significant increase in the protein expression of the HPC-specific protein CK19 (0.901±0.072 vs 0.686±0.097, t=-3.093, P＜0.05). Western blot showed that compared with the D12 control group, the D12 MC group had a significant increase in the protein expression of the HPC-related protein CK19 (1.922±0.103 vs 1.448±0.012, t =-7.881, P ＜005), as well as a significant increase in the protein expression of the autophagy-related protein LC3 (1.392±0.042 vs 1.101±0048, t =-5.978, P＜005). ConclusionMCs can promote the differentiation of mouse iPSCs into HPCs, possibly by increasing the autophagy level of HPCs.

Capacidad multipotencial en células madre mesenquimales derivadas de la placenta humana en Panamá / Multipotent capacity of human placenta­derived mesenchymal stem cells in Panama

Introducción: Las células madre mesenquimales (MSCs) tienen la capacidad única de auto­renovación y pluripotencia con la cual apoyan en la regeneración de tejidos en or­ ganismos vivos. El mayor potencial terapéutico de las MSCs derivadas de la placenta (PDMSCs) humana, como fuente más joven de MSCs, estimula la búsqueda de las me­ jores condiciones de cultivo que preserven su capacidad de proliferar y diferenciarse. Sin embargo, estudios relacionados a la caracterización de la multipotencialidad de las PDMSCs durante periodos prolongados de cultivo, no han sido reportados en Panamá. Por lo tanto, el objetivo de este estudio fue el de implementar un proceso de aislamiento y cultivo que preserve las propiedades multipotentes en PDMSCs. Materiales y Méto­ dos: Placentas humanas a término completo fueron obtenidas para el aislamiento de las MSCs. Las PDMSCs fueron caracterizadas según su morfología, expresión positiva de marcadores CD90, CD73, CD105, y capacidad de proliferación y diferenciación a linajes mesodérmicos. Resultados: Se ha demostrado la obtención de poblaciones de PDMSCs con morfología fibroblástica, adherencia plástica, expresión positiva de los marcadores CD90, CD73 y CD105, y capacidad de diferenciación osteogénica, adi­ pogénica y condrogénica. Posterior al aislamiento y crio­preservación, las PDMSCs mantuvieron una viabilidad mayor de 95%, una tasa de proliferación por más de 40 días en cultivo, y la expresión positiva de los marcadores CD90, CD73, y CD105 al pasaje 16. Conclusión: Nuestros resultados demuestran una metodología eficiente para obten­ ción y cultivo de PDMSCs que mantienen sus características multipotentes durante períodos prolongados de cultivo, abriendo el camino para futuras terapias celulares

ntroduction: Human mesenchymal stem cells are (MSCs) unique in their pluripotency and their ability to self­renew in order to support tissue regeneration in living organisms. The increased therapeutic potential of PDMSCs as a pool of younger MSCs with a vast capacity for expansion, minor predisposition for tumor formation or immune reactions spurs the search for the best culture conditions to preserve their ability to differentiate and proliferate. However, studies regarding characterization of multipotent isolated PDMSCs during prolonged periods of culture has not been reported thus far in Panama. Therefore, in this study we seek to implement isolation and culture procedures that pre­ serve multipotent characteristics in PDMSCs. Materials and Methods: Full­term human placentas were obtained for the isolation of MSCs. PDMSCs where characterized based on their morphology, positive expression of CD90, CD73, and CD105, and their ability to proliferate and differentiate to mesodermal lineages. Results: It was demonstrated that our isolated PDMSCs presented the MSC characteristics of fibroblastic morphology, plastic adherence, positive expression of CD90, CD73, and CD105 markers, as well as osteogenesis, adipogenesis, and osteogenic differentiation ability. When PDMSCs were cultured after isolation or cryopreservation, viability was maintained above 95%, with their proliferation rate maintained after 40 days, and positive expression of CD90, CD73, and CD105 markers kept after 16 passages. Conclusion: Taken together, our results de­ monstrated a methodology to obtain successful source of isolated human PDMSCs that kept their multipotent properties over time, opening the path for future cellular therapies.

Induced pluripotent stem cells: advances in research and application in dermatology / 中华皮肤科杂志

Induced pluripotent stem cells (iPSCs) are a kind of stem cells induced from somatic cells by reprogramming with some specific pluripotent transcription factors.Compared with traditional stem cells,iPSCs have many advantages,including a wide range of sources,the ability of totipotent differentiation,and less ethical and sourcing issues associated with embryonic stem cells which have been debated for a long time.In dermatology,iPSCs can be used to make regenerative skin,establish models of skin diseases,provide a new platform for studying the pathogenesis of skin diseases,and perform cell treatment or gene modification treatment for specific skin diseases.At present,the application value of iPSCs has been verified among animal models,and clinical trials are also under way.

Molecular and cellular mechanisms underlying the Stem Cell Educator therapy for the clinical treatment of diabetes / 中国医师杂志

Multiple immune dysfunctions and shortage of islet beta cells are two key issues for type 1 (T1D) and type 2 (T2D) diabetes.International multi-center clinical studies and basic research have demonstrated the safety and clinical efficacy of Stem Cell Educator therapy for the treatment of T1D and T2D.CB-SC display multiple immune modulations on T cells,regulatory T cells (Tregs),and monocytes through various molecular mechanisms,such as cell-cell contacting,releasing soluble factors,and correcting the autoimmune memory.Recently,we found that platelet-releasing mitochondria exhibit the immune modulation and can migrate to pancreatic islets and be taken up by islet beta cells,leading to the proliferation and enhancement of islet beta cell functions.These findings reveal new mechanisms underlying Stem Cell Educator therapy and open up new avenues to improve the treatment of diabetes in clinics.

GaAlAs laser pretreatment enhances the function of ADSCs in photoaging of the skin in rats / 中华医学美学美容杂志

Objective To investigate the effect of adipose-derived stem cells (ADSCs) on pho toaging skin after laser pretreatment with GaAlAs.Methods ADSCs were isolated from healthy wistar rats,ADSCs were isolated and cultured to establish an adipose-derived stem cell culture system.ADSCs were pretreated with GaAlAs laser at a wavelength of 650 nm 4 J/cm2.A rat model of pho toaging aging was established.Different doses of ADSCs and low energy laser ADSCs were pretreated with ADSCs for the treatment of photoaging skin,and the morphological changes of epidermis and dermis were observed before and after treatment with low energy laser pretreatment.Results When the concentration of ADSCs was 103/100 μl,there was no significant difference in epidermal thickness and dermal thickness between ADSCs treated group and GaAlAs pretreatment group (P＞0.05).The thickness of epidermis in the GaAlAs pretreatment group was significantly lower than that in the ADSCs group (P＜0.05) at 104/100 μl.When the concentration of ADSCs was 5 × 104/100 μl,the epidermal thickness of the GaAlAs pretreatment group decreased significantly and the thickness of the dermis increased significantly,which was significantly different from that of the ADSCs group (P ＜ 0.05).Conclusions GaAlAs laser pretreatment can enhance ADSCs anti-skin photoaging ability.

Advances in research and application of liver organoids / 临床肝胆病杂志

Liver diseases significantly affect human health. Classic research models cannot maintain hepatocyte polarity for a long time and have a different genetic background from normal human hepatocytes, which limits the research on the physiological functions of hepatocytes and the mechanisms of related diseases. The latest liver organoid technology can obtain liver organoids with partial liver structure by reprogrammed induced pluripotent stem cells or in vitro culture of liver biopsy tissue and maintain long-term proliferation and genetic stability, and therefore, it is expected to become a powerful tool for disease modelling, drug screening, and cell therapy. This article mainly reviews the research and application of liver organoids in recent years.

Comparison of the characteristics of canine adipose tissue-derived mesenchymal stem cells extracted from different sites and at different passage numbers

Mesenchymal stem cells (MSCs) have desirable characteristics for use in therapy in animal models and veterinary medicine, due to their capacity of inducing tissue regeneration and immunomodulation. The objective of this study was to evaluate the differences between canine adipose tissue-derived MSCs (AD-MSCs) extracted from subcutaneous (Sc) and visceral (Vs) sites. Surface antigenic markers, in vitro differentiation, and mineralized matrix quantification of AD-MSCs at different passages (P₄, P₆, and P₈) were studied. Immunophenotypic analysis showed that AD-MSCs from both sites were CD44+, CD90+, and CD45−. Moreover, they were able, in vitro, to differentiate into fat, cartilage, and bone. Sc-AD-MSCs preserve in vitro multipotentiality up to P₈, but Vs-AD-MSCs only tri-differentiated up to P₄. In addition, compared to Vs-AD-MSCs, Sc-AD-MSCs had greater capacity for in vitro mineralized matrix synthesis. In conclusion, Sc-AD-MSCs have advantages over Vs-AD-MSCs, as Sc AD-MSCs preserve multipotentiality during a greater number of passages, have more osteogenic potential, and require less invasive extraction.

Advances in the research of the application of induced pluripotent stem cells in tissue engineering skin as seed cells / 中华烧伤杂志

Tissue engineering skin has a wide application prospect on the clinical treatment of all sorts of skin defect, especially large area burn. The shortage of seed cells and their function improvement are the main problems in this field. The existing seed cells of tissue engineering skin are difficult to meet the needs of clinical application due to the limitations of acquisition, proliferation, and aging. Subsequently, the generation of induced pluripotent stem cells (iPSCs) provides a safe and efficient cell source for tissue engineering skin. Our article focuses on the origin of iPSCs and its characteristics of differentiating into keratinocytes, fibroblasts, melanocytes, vascular smooth muscle cells, nerve cells, and hair follicle, and discusses the main problems and prospects of iPSCs in establishment of tissue engineering skin and application in wound repair.

Leukemia propagating cells in Philadelphia chromosome-positive ALL: a resistant phenotype with an adverse prognosis

BACKGROUND: Targeted therapy has revolutionized the management of Philadelphia chromosome-positive (Ph+) acute lymphoblastic leukemia (ALL); however, relapse still occurs because of the presence of quiescent stem cells, termed leukemia propagating cells (LPCs). This study aimed to assess the phenotypic diversity of LPCs in adult patients with Ph+ B-Acute ALL (B-ALL) and to assess its prognostic impact. METHODS: Seventy adults with newly diagnosed Ph+ B-ALL were recruited at the Mansoura Oncology Center. Multiparameter flow cytometry studies of mononuclear blast cells for cluster of differentiation (CD)34, CD38, and CD58 were performed. RESULTS: Seventeen patients had blasts with the pattern of LPCs (CD34+CD38−CD58−), while 53 cases had other diverse phenotypic patterns. The rate of complete response was significantly lower in patients with the LPC phenotype (47% vs. 81%, P=0.006). The median time to achieve a complete response was prolonged in patients with the CD34+CD38−CD58− phenotype (48 vs. 32 days, P=0.016). The three-year overall survival was significantly lower in patients with the CD34+CD38−CD58− phenotype (37% vs. 55% respectively, P=0.028). Multivariate analysis showed that the CD34+CD38− CD58− phenotype was an independent risk factor for overall survival. CONCLUSION: The presence of CD34+CD38−CD58− LPCs at diagnosis allows rapid identification of higher risk patients. Risk stratification of these patients is needed to further guide therapy and develop effective LPCs-targeted therapy to improve treatment outcome.

The Therapeutic Effect of Human Embryonic Stem Cell-Derived Multipotent Mesenchymal Stem Cells on Chemical-Induced Cystitis in Rats / 대한배뇨장애요실금학회지

PURPOSE: To evaluate the therapeutic effect of human embryonic stem cell (hESC)-derived multipotent mesenchymal stem cells (M-MSCs) on ketamine-induced cystitis (KC) in rats. METHODS: To induce KC, 10-week-old female rats were injected with 25-mg/kg ketamine hydrochloride twice weekly for 12 weeks. In the sham group, phosphate buffered saline (PBS) was injected instead of ketamine. One week after the final injection of ketamine, the indicated doses (0.25, 0.5, and 1×106 cells) of M-MSCs (KC+M-MSC group) or PBS vehicle (KC group) were directly injected into the bladder wall. One week after M-MSC injection, the therapeutic outcomes were evaluated via cystometry, histological analyses, and measurement of gene expression. Next, we compared the efficacy of M-MSCs at a low dose (1×105 cells) to that of an identical dose of adult bone marrow (BM)-derived MSCs. RESULTS: Rats in the KC group exhibited increased voiding frequency and reduced bladder capacity compared to rats of the sham group. However, these parameters recovered after transplantation of M-MSCs at all doses tested. KC bladders exhibited markedly increased mast cell infiltration, apoptosis, and tissue fibrosis. Administration of M-MSCs significantly reversed these characteristic histological alterations. Gene expression analyses indicated that several genes associated with tissue fibrosis were markedly upregulated in KC bladders. However the expression of these genes was significantly suppressed by the administration of M-MSCs. Importantly, M-MSCs ameliorated bladder deterioration in KC rats after injection of a low dose (1×105) of cells, at which point BM-derived MSCs did not substantially improve bladder function. CONCLUSIONS: This study demonstrates for the first time the therapeutic efficacy of hESC-derived M-MSCs on KC in rats. M-MSCs restored bladder function more effectively than did BM-derived MSCs, protecting against abnormal changes including mast cell infiltration, apoptosis and fibrotic damage.

Current and Future Perspectives of Stem Cell Therapy in Dermatology

Stem cells are undifferentiated cells capable of generating, sustaining, and replacing terminally differentiated cells and tissues. They can be isolated from embryonic as well as almost all adult tissues including skin, but are also generated through genetic reprogramming of differentiated cells. Preclinical and clinical research has recently tremendously improved stem cell therapy, being a promising treatment option for various diseases in which current medical therapies fail to cure, prevent progression or relieve symptoms. With the main goal of regeneration or sustained genetic correction of damaged tissue, advanced tissue-engineering techniques are especially applicable for many dermatological diseases including wound healing, genodermatoses (like the severe blistering disorder epidermolysis bullosa) and chronic (auto-)inflammatory diseases. This review summarizes general aspects as well as current and future perspectives of stem cell therapy in dermatology.

Comparative study of proliferative and periodontal differentiation propensity of induced pluripotent stem cells at different passages / 北京大学学报(医学版)

Objective:To compare the proliferative and periodontal specific differentiation abilities of induced pluripotent stem cells (iPSCs) at different passages,and to investigate whether long term culturing would have a negative influence on their proliferation and specific differentiation capacity,thus providing a theoretical basis for further in-depth research on periodontal regeneration and the possible clinical applications of iPSCs.Methods:IPSCs derived from human gingival fibroblasts at passages 5,10,15 and 20 were recovered and cultured in vitro.Their morphology and proliferation rates were observed respectively.We further induced the iPSCs at different passages toward periodontal tissue under the treatment of growth/differentiation factor-5 (GDF-5) for 14 days through the EB routine,then compared the periodontal differentiation propensities between the different passages of iPSCs by detecting their calcified nodules formation by Alizarin red staining and assaying their relative periodontal tissue related marker expressions by qRT-PCR and immunofluorescence staining,including bone related markers:osteocalcin (OCN),bone sialoprotein (BSP);periodontal ligament related markers:periostin,vimentin;and cementum related markers:cementum attachment protein (CAP),cementum protein 1 (CEMP1).The untreated spontaneous differentiation groups were set as negative controls respectively.Results:iPSCs at different passages all showed a high proliferative capacity when cultured in vitro and turned into a spindlelike shape similar to fibroblasts upon periodontal specific differentiation.All iPSCs formed typical calcified nodules upon GDF-5 induction by Alizarin red staining in comparison to their untreated controls.The relative calcium deposition at all passages had been significantly upgraded under the treatment of GDF-5 (P5:t =2.125,P =0.003;P10:t =2.246,P =0.021;P15:t =3.754,P =0.004;P20:t =3.933,P =0.002),but no significant difference in their calcium deposition were detected within passages 5,10,15 and 20 (periodontal differentiation:F =2.365,P =0.109;spontaneously differentiation:F =2.901,P =0.067).Periodontal tissue related marker expressions of iPSCs at all passages had also been significandy upgraded under the treatment of GDF-5 (P ＜ 0.05),but still,no significant difference in their expression levels of periodontal tissue related proteins were detected within passages (BSP:F =0.926 7,P=0.450;vimentin:F=0.917 1,P=0.455;CEMP1:F=2.129,P=0.1367).Conclusion:Our results preliminarily confirmed that long term culturing won't influence the proliferation capacity and periodontal specific differentiation propensity of iPSCs,as they can still proliferate and differentiate toward periodontal cells with high efficiency upon growth factor induction after continuous passaging.Therefore,iPSCs could be recognized as a promising cell source for future possible application in periodontal tissue regeneration.

Application of swine iPS cell transplantation in porcine myocardial infarction model / 中国医师杂志

Objective To observe the therapeutic effect of porcine myocardium against porcine myocardial infarction (MI) by porcine myocardial infarction model with connexin-43/zonula occludens-1 (CX43/ZO-1) double gene induced pluripotent stem cells (iPS) transplantation.Methods Porcine myocardial infarction model was established by balloon occlusion.One week later,1640 culture medium,CX43 gene ips cells and CX43/ZO-1 gene ips cells were injected into the different coronary arteries.After 3 months,electron microscopy and color Doppler ultrasonography were used to evaluate the curative effect.Results Compared to ips group of CX43 and 1640 culture group,in CX43/ZO-1 gene ips group,electron microscope showed a large number of island-like cardiomyocytes,blood vessels and intact intercalated discs in the infarct area and normal myocardial junction area structure,color Doppler echocardiography showed left ventricular end diastolic diameter reduced,and ejection fraction was significantly increased (P ＜ 0.05).Conclusions Transfection of CX43/ZO-1 double gene ips cells can promote cardiomyocyte regeneration and cardiac function recovery in porcine myocardial infarction model.

Immune modulation by mouse lymphocytes co-culturedwith cord blood-derived multipotent stem cells in Alzheimer's disease model mice / 基础医学与临床

Objective To investigate the immune modulation effect of lymphocytes co-cultured with human cord blood derived-multipotent stem cells(CB-SCs) and to explore their therapeutic potential for Alzheimer's Disease (AD) model mice.Methods CB-SCs were isolated from human cord blood.Lymphocytes were isolated from spleens of AD model mice.The lymphocytes were co-cultured with CB-SCs or cultured alone for 72 h.AD model mice were divided into experimental group and control group randomly,and then the experimental group mice were administrated with lymphocytes co-cultured with CB-SCs and control group were administrated with lymphocytes cultured alone by caudal vein injection.Then,the behavior experiment was carried out.The CD4+CD25+Foxp3+Tregswere detected by flow cytometry.The protein expression of TNF-αand IL-10 in peripheral blood was detected by ELISA,The mRNA expression of TNF-α and IL-10 in brain tissue was detected by PCR.The amyloid-β(Aβ)plaques were detected by immunohistochemistry.Results 1)There was spatial learning and memory improvement on experimental group.2)The Aβ plaques of experimental group decreased.3)The percentage of CD4+CD25+Foxp3+Tregs in peripheral blood and IL-10 level in plasma were higher in experimental group(P<0.05).The pro-inflammatory cytokines,TNF-α level in plasma of experimental group was lower than that in the control group(P<0.001).4)The mRNA expression of IL-10 in brain was higher in experimental group as compared to those in the control group(P<0.05),and the mRNA expression of TNF-α of experimental group was lower than that in the control group(P<0.05).Conclusions The lymphocytes co-cultured with CB-SCs have immunotherapeutic effect in AD model mice,which is mainly displayed with increased proportion of Tregs and enhanced anti-inflammatory function of Tregs.

Generation of pluripotent stem cells using non-integrated approach in human azoospermia patients / 天津医药

Objective To establish induced pluripotent stem cells (iPSCs) in patients with azoospermia by non-integrated approach. Methods Using the commercially available serum-free medium (TeSR?2) and embryonic stem cell culture medium (Stem Adhere? Defined Matrix) to define the culture system, the iPSCs were established by using non-integrated Sendai virus infection in peripheral blood mononuclear cells of azoospermia patients. The immunofluorescence, karyotype analysis, embryoid body differentiation and teratoma formation were used to identify pluripotency, karyotype and differentiation ability of iPSCs. Results The established iPSCs showed the characteristics of human embryonic stem cells. Immunofluorescence analysis showed that octamer-binding transcription factor 4 (OCT4), SRY-related-box protein-2 (SOX2), stage-specific embryonic antigen-4 (SSEA-4) and tumor rejection antigen-1-60 (TRA-1-60) were positive for the expression of stem cell pluripotency markers. Karyotype analysis showed that they had normal karyotype. In addition, embryoid body and teratoma tests showed that the iPSCs had the ability to differentiate into three germ layers in vitro and in vivo. Conclusion The induction of pluripotent stem cell line is successfully constructed by non-integrated approach in azoospermia patients.

Umbilical cord blood pluripotent stem cell immune education treatment for juvenile type 1 diabetes / 中国医师杂志

Type 1 diabetes is a autoimmune disease that occurs under the influence of both genetic predisposition and environmental factors,mediated mainly by T lymphocytes with various kinds of other involved immune cells.The autoimmune attacks eventually lead to the islet beta cell destruction and insulin insufficiency.Multiple fundamental studies and clinical trials have revealed that umbilical cord blood pluripotent stem cells have the potential to help restore immune balance,induce immune tolerance,stop the autoimmune attacks against beta cells,and promote beta cell regeneration in type 1 diabetes mellitus (T1DM),by means of cell to cell contact and soluble cytokine secretion.For the past few years,the National Clinical Research Center for Metabolic Diseases of Second Xiangya Hospital has been leading the research on stem cell education therapy and has performed the therapy for 35 juvenile type 1 diabetes patients from China and foreign countries,introducing a novel treatment for T1DM.

Extensive characterization of feline intra-abdominal adipose-derived mesenchymal stem cells

Mesenchymal stem cells (MSCs) isolated from various tissues have been well characterized for therapeutic application to clinical diseases. However, in contrast to MSCs from other animal species, the characteristics of feline MSCs have not been fully documented. In this study, we conducted extensive characterization of feline adipose tissue-derived MSCs (fAD-MSCs). Study fAD-MSCs were individually isolated from the intra-abdominal adipose tissues of six felines. The expression levels of cell surface markers and pluripotent markers were evaluated. Next, proliferation capacity was analyzed by performing cumulative population doubling level (CPDL) and doubling time (DT) calculation assays. Differentiation potentials of fAD-MSCs into mesodermal cell lineages were analyzed by examining specific staining and molecular markers. All fAD-MSCs positively expressed cell surface markers such as CD29, CD44, CD90, CD105, CD166, and MHC-I, while CD14, CD34, CD45, and CD73 were negatively expressed. The CPDL of the fAD-MSCs was maintained until passage 5 to 6 (P5 to P6), whereas DT increased after P3 to P4. Also, stem cell-specific pluripotent markers (Oct3/4, Nanog, and SSEA-4) were detected. Importantly, all fAD-MSCs demonstrated mesodermal differentiation capacity. These results suggest that fully characterized fAD-MSCs could be beneficial when considering the use of these cells in feline disease research.