Cobalt Chloride as a Hypoxia Mimicking Agent Induced HIF-1α and mTOR Expressions of Human Umbilical Cord Mesenchymal Stem Cells

ABSTRACT Objective: To assess the effects of cobalt chloride (CoCl2) as a hypoxia mimicking agent on human umbilical cord mesenchymal stem cells (hUCMSCs) expression of HIF-1α and mTOR for use in regenerative dentistry. Material and Methods: Human umbilical cord mesenchymal stem cells were isolated and then cultured. The characteristics of stemness were screened and confirmed by flow cytometry. The experiment was conducted on hypoxia (H) and normoxia (N) groups. Each group was divided and incubated into 24-, 48-, and 72-hours observations. Hypoxic treatment was performed using 100 µM CoCl2 on 5th passage cells in a conventional incubator (37°C; 5CO2). Then, immunofluorescence of HIF-1α and mTOR was done. Data was analyzed statistically using One-way ANOVA and Tukey's HSD. Results: Significant differences were found between normoxic and hypoxic groups on HIF-1α (p=0.015) and mTOR (p=0.000) expressions. The highest HIF-1α expression was found at 48 hours in the hypoxia group, while for mTOR at 24 hours in the hypoxia group. Conclusion: Hypoxia using cobalt chloride was able to increase human umbilical cord mesenchymal stem cells expression of HIF-1α and mTOR.

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.

HucMSC-Ex alleviates inflammatory bowel disease via the lnc78583-mediated miR3202/HOXB13 pathway / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

As a group of nonspecific inflammatory diseases affecting the intestine, inflammatory bowel disease (IBD) exhibits the characteristics of chronic recurring inflammation, and was proven to be increasing in incidence (Kaplan, 2015). IBD induced by genetic background, environmental changes, immune functions, microbial composition, and toxin exposures (Sasson et al., 2021) primarily includes ulcerative colitis (UC) and Crohn's disease (CD) with complicated clinical symptoms featured by abdominal pain, diarrhea, and even blood in stools (Fan et al., 2021; Huang et al., 2021). UC is mainly limited to the rectum and the colon, while CD usually impacts the terminal ileum and colon in a discontinuous manner (Ordás et al., 2012; Panés and Rimola, 2017). In recent years, many studies have suggested the lack of effective measures in the diagnosis and treatment of IBD, prompting an urgent need for new strategies to understand the mechanisms of and offer promising therapies for IBD.

Expression of Twist1, SIRT1, FGF2 and TGF-β3 genes and its regulatory effect on the proliferation of placenta, umbilical cord and dental pulp mesenchymal stem cells / 中华医学遗传学杂志

OBJECTIVE@#To compare the mRNA level of cell proliferation-related genes Twist1, SIRT1, FGF2 and TGF-β3 in placenta mesenchymal stem cells (PA-MSCs), umbilical cord mensenchymals (UC-MSCs) and dental pulp mesenchymal stem cells (DP-MSCs).@*METHODS@#The morphology of various passages of PA-MSCs, UC-MSCs and DP-MSCs were observed by microscopy. Proliferation and promoting ability of the three cell lines were detected with the MTT method. Real-time PCR (RT-PCR) was used to determine the mRNA levels of Twist1, SIRT1, FGF2, TGF-β3.@*RESULTS@#The morphology of UC-MSCs and DP-MSCs was different from that of PA-MSCs. Proliferation ability and promoting ability of the PA-MSCs was superior to that of UC-MSCs and DP-MSCs. In PA-MSCs, expression level of Twist1 and TGF-β3 was the highest and FGF2 was the lowest. SIRT1 was highly expressed in UC-MSCs. With the cell subcultured, different expression levels of Twist1, SIRT1, FGF2, TGF-β3 was observed in PA-MSCs, UC-MSCs and DP-MSCs.@*CONCLUSION@#Up-regulated expression of the Twist1, SIRT1 and TGF-β3 genes can promote proliferation of PA-MSCs, UC-MSCs and DP-MSCs, whilst TGF-β3 may inhibit these. The regulatory effect of Twist1, SIRT1, FGF2 and TGF-β3 genes on PA-MSCs, UC-MSCs and DP-MSCs are different.

Efficient Isolation and Long-term Red Fluorescent Nanodia-mond Labeling of Umbilical Cord Mesenchymal Stem Cells for the Effective Differentiation into Hepatocyte-like Cells

Abstract Fluorescent nanodiamond (FND) has been used for long-term cell labeling and in vivo cell tracking because they have good at photostability and biocompatibility. In this study, we evaluate the effect of fluorescent nanodiamond labeling on in vitro culture and differentiation of human umbilical cord mesenchymal stem cells (hUCMSCs) into hepatocyte-like cells (HLCs). For hepatic differentiation of hUCMSCs, cells were induced with human hepatocyte growth factor, nicotinamide and Dexamethasone. FND was supplied in two experimental groups with 20 μg/mL and 100 μg/mL in 2 hours. The cell was assessed for FND uptake by laser scan microscopy and flow cytometry methods. The effect of FND on hUCMSCs was evaluated by the cell viability and growth assays as well as the differentiation throughout of morphology alterations or gene expression of anfa-fetoprotein, albumin, and hepatocyte nuclear factor 4α. The results showed that the labeling of hUCMSCs is efficient and easy and there was significant cellular uptake of FND. We did not observe any negative impacts of FND to the cell viability and growth. FND can be utilized for the long-term labeling and tracking of hUCSCs and HLCs in vivo studies.

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.

Angiopoietin-1 Modified Human Umbilical Cord Mesenchymal Stem Cell Therapy for Endotoxin-Induced Acute Lung Injury in Rats

PURPOSE: Angiopoietin-1 (Ang1) is a critical factor for vascular stabilization and endothelial survival via inhibition of endothelial permeability and leukocyte- endothelium interactions. Hence, we hypothesized that treatment with umbilical cord mesenchymal stem cells (UCMSCs) carrying the Ang1 gene (UCMSCs-Ang1) might be a potential approach for acute lung injury (ALI) induced by lipopolysaccharide (LPS). MATERIALS AND METHODS: UCMSCs with or without transfection with the human Ang1 gene were delivered intravenously into rats one hour after intra-abdominal instillation of LPS to induce ALI. After the rats were sacrificed at 6 hours, 24 hours, 48 hours, 8 days, and 15 days post-injection of LPS, the serum, the lung tissues, and bronchoalveolar lavage fluid (BALF) were harvested for analysis, respectively. RESULTS: Administration of fluorescence microscope confirmed the increased presence of UCMSCs in the injured lungs. The evaluation of UCMSCs and UCMSCs-Ang1 actions revealed that Ang1 overexpression further decreased the levels of the pro-inflammatory cytokines TNF-α, TGF-β1, and IL-6 and increased the expression of the anti-inflammatory cytokine IL-10 in the injured lungs. This synergy caused a substantial decrease in lung airspace inflammation and vascular leakage, characterized by significant reductions in wet/dry ratio, differential neutrophil counts, myeloperoxidase activity, and BALF. The rats treated by UCMSCs-Ang1 showed improved survival and lower ALI scores. CONCLUSION: UCMSCs-Ang1 could improve both systemic inflammation and alveolar permeability in ALI. UC-derived MSCs-based Ang1 gene therapy may be developed as a potential novel strategy for the treatment of ALI.

Umbilical cord mesenchymal stem cells in neurological disorders: A clinical study.

We investigated the intrathecally administrated unbilical cord mesenchymal stem cells (UC-MSCs) by lumbar puncture and assessed the technical difficulties and effects in various neurological conditions. One hundred patients underwent subarachnoid placement of UC-MSCs between December 2006 and May 2010 in the Affiliated Hospital of Medicine. Technical difficulties in patients in the form of localization of subarachnoid space, number of attempts, and post-procedural complications were evaluated. Functional evaluation was done using Hauser Ambulation Index (HAI) by the stem cell transplant team on a regular basis. All patients were followed-up for more than 1 yr after the treatment. Clinical symptoms, related biochemical index and photographic examinations were observed regularly. We encountered technical difficulties in 31 patients (31%) in the form of general anesthesia supplementation and difficulty localizing the lumbar space. Side effects (headache, low-grade fever, low back pain and lower limb pain) were observed in 22 (22%) patients, which were treated with symptomatic therapy within 48 h. One year after the treatment, functional indices improved in 47 patients (47%): 12 patients with spinal cord injury, 11 patients with cerebral palsy, 9 patients with post-traumatic brain syndrome, 9 patients with post-brain infarction syndrome, 3 patients with spinocerebellar ataxias, and 3 patients with motor neuron disease. In conclusion,intrathecal administration of UC-MSCs is a safe and effective way to treat neurological disorders. Our encouraging results of intrathecal administration of UC-MSCs indicate the potential of restoration of lost tissue and improvement of function in patients with profound neurological defects and inefficient conventional cure. These data support expanded double-blind, placebo-controlled studies for this treatment modality.

Effects of Human Umbilical Cord Mesenchymal Stem Cells on Renal Ischaemia-reperfusion Injury in Rats

Objective This study aims to observe the function of umbilical cord-mesenchymal stem cells (UC-MSCs) labelled with enhanced green fluorescent protein (eGFP) in the repair of renal ischaemia-reperfusion (I/R) injury, to determine the effects on inflammatory cascade in an established rat model and to explore possible pathogenesis. Materials and Methods Sixty rats were randomly divided into three groups: the sham-operated, I/R and UC-MSC treatment groups. All rats underwent right nephrectomy. Ischaemia was induced in the left kidney by occlusion of the renal artery and vein for 1hour, followed by reperfusion for 24 hours or 48 hours. Kidney samples were collected to observe morphological changes. Immunohistochemistry was performed to assess the expression of intercellular adhesion molecule 1 (ICAM-1) in the renal tissue sample, as well as the number of infiltrating polymorphonuclear neutrophils (PMNLs) and UC-MSCs with positive eGFP. Results Renal histopathological damages and the expression of ICAM-1 and PMNL increased significantly in the I/R group compared with those in the sham-operated group, whereas the damages were less conspicuous in the UC-MSC treatment group. Conclusions Renal ICAM-1, which mediated PMNL infiltration and contributed to renal damage, was significantly up-regulated in the I/R group. UC-MSCs were identified to inhibit these pathological processes and protect the kidney from I/R injury. .

[Generation of induced pluripotent stem cells from mesenchymal stem cells of human umbilical cord vein]

The aim of this study was to generate induced pluripotent stem cells from human umbilical cord vein mesenchymal stem cells by use of plasmid vector. In this experimental study using type IV collagenase enzyme, we extracted and cultured mesenchymal stem cells obtained from human umbilical cord vein wall.Usingelectroporation method, these cells were transfected with plasmid vector which carried selfrenewal transcription factors of OCT4 and SOX2.After 9 days we observed the induced pluripotent stem cells like colonies. The nature of thesecells were evaluated byimmunocytochemistry and alkalin phosphatase activity of embryonic stem cells. Immunocytochemical analysis showed that these cells express the pluripotency markerssuch as OCT4, SSEA4, TRA1-60, TRA1-81, and also hadalkalin Phosphatase activity. This study revealed that transient expression of self renewal genes of OCT4 and SOX2 could lead to development of induced pluripotent stem cells like colonies from umbilical cord vein mesenchymal stem cells

Validación del sistema UMELISA HBsAg PLUS utilizando muestras de suero de cordón umbilical / Validation of UMELISA HBsAg PLUS system based on umbilical cord serum samples

Introducción: la selección y el desarrollo de métodos analíticos siempre ha sido un tema importante para los laboratorios de ensayo. Para adoptar un método se hace necesario validarlo, porque el objetivo de la validación es probar la aptitud de los métodos, así como la competencia del laboratorio para realizar determinado ensayo. El sistema UMELISA HBsAg PLUS no contempla dentro de su aplicación el empleo de muestras de suero obtenidas a partir de sangre del cordón umbilical, sin embargo, con este tipo de muestra se certifica el uso de la placenta como materia prima. Objetivo: teniendo en cuenta la necesidad de demostrar que el cambio no afecta los parámetros de desempeño del sistema, se hizo su validación para muestras de suero de cordón umbilical. Métodos: se emplearon tres paneles de muestras en los estudios de especificidad, concordancia con el sistema de referencia Hepanostika HBsAg Uni-Form II, robustez, precisión a dos niveles de repetibilidad (intraensayos) y precisión intermedia (interensayos), y límite de detección. Resultados: la validación del sistema mostró los siguientes indicadores: especificidad y concordancia del 100 porciento, sistema robusto frente a los cambios de temperatura entre 35 y 38 °C, elevada precisión intraensayos (4,65 porciento) e interensayos (CV menores que 20 porciento), límite de detección correspondiente a una dilución de 1/10 000. Conclusiones: los resultados demostraron que el sistema UMELISA HBsAg PLUS se puede emplear para la detección de HBsAg en muestras de suero de cordón umbilical

Introduction: the selection and the development of analytical methods have always been an important issue for testing laboratories. The adoption of a method requires validation because the objective is to prove the capability of a method as well as the competencies of the laboratory to make an specific assay. The range of applications of UMELISA HbsAg PLUS system does not include the use of serum samples taken from the umbilical cord blood; however the use of placenta as raw material is certified with this kind of sample. Objective: to validate this system for umbilical cord serum sample taking into account the need of proving that this change does not affect the performance parameters of the system. Methods: three panels of samples were used to study specificity, agreement with the reference system Hepanostika HbsAg Uni-Form II, robustness, precision at two levels of repeatibility (intrassay) and intermediate precision (interassay) and detection limit. Results: validation of the system showed the following indicators: specificity and agreement of 100 percent, robust system against temperatures changes between 35 and 38 °C, high intraassay precision (4.65 percent) and interassays (VC less than 20 percent), detection limit corresponding to 1/10 000 dilution rate. Conclusions: obtained results showed that UMELISA HbsAg PLUS system may be used to detect HBsAg in umbilical cord serum samples

Eficacia del cordón umbilical humano como sustrato para detección de anticuerpos anti endomisio / Efficiency of human umbilical cord as substrate or detection of antiemdomysium antibodies / Aeficácia do cordão umbilical humano como substrato para a detenção de anticorpos anti-endomísio

Los anticuerpos anti endomisio IgA (EMA) están dirigidos hacia antígenos del tejido conectivo que rodea a las fibras del músculo liso. El objetivo de este trabajo fue evaluar la eficacia del cordón umbilical humano (CUH) como sustrato para detectar EMA mediante inmunofluorescencia indirecta y compararlo con una de las metodologías disponibles comercialmente, la cual utiliza como sustrato esófago de mono. Se obtuvieron 100 sueros de pacientes con diagnóstico de enfermedad celíaca y 50 sueros de pacientes clínicamente sanos con biopsia de mucosa intestinal normal, los cuales realizaron su consulta y atención en el Hospital Privado Centro Médico de Córdoba, en un periodo de tiempo comprendido entre los años 2006 y 2009. Los resultados obtenidos mostraron una "muy buena" concordancia entre ambos métodos. Se estimó para el método que utiliza CUH una sensibilidad y especificidad de 98% (93-99%) y 100% (93-100%) respectivamente con una eficacia del 99%. De acuerdo con lo anterior se concluye que utilizar CUH como sustrato para evaluar la presencia de EMA es confiable y efectivo para detectar pacientes con enfermedad celíaca no tratada.

The antiendomysium antibodies (EMA) are directed toward antigens of connective tissue that surrounds the smooth muscle fibers. The aim of this study was to evaluate the efficiency of human umbilical cord (HUC) as substrate to detect EMA by indirect immunofluorescence and to compare it with one of the commercially available methodologies which use monkey esophagus as substrate. Serum samples obtained from 100 patients with celiac disease diagnosis and 50 healthy controls with normal intestinal mucosa were evaluated. Patients were treated at the Hospital Privado Centro Médico de Córdoba over a period of time between 2006 and 2009. The results showed an "almost perfect" concordance between both methods. The calculated sensitivity and specificity for HUC was 98% (93-99%) and 100% (93-100%) respectively, with an efficiency of 99%. This results indicate that the use of HUC as substrate to evaluate the presence of EMA is reliable and effective for the detection of patients with untreated celiac disease.

Os anticorpos anti-endomísio IgA (EMA) são direcionados contra os antígenos do tecido conectivo que cercam as fibras do músculo liso. O objetivo deste trabalho foi avaliar a eficácia do cordão umbilical humano (CUH) como substrato para detectar EMA através da imunofluorescência indireta e compará-lo com uma das metodologias disponíveis comercialmente, a qual utiliza como substrato esôfago de macaco. Foram obtidos 100 soros de pacientes com diagnóstico de doença celíaca e 50 soros de pacientes clinicamente saudáveis com biópsia de mucosa intestinal normal, os quais realizaram sua consulta e atendimento no Hospital Privado Centro Médico de Córdoba, em um período de tempo compreendido entre os anos 2006 e 2009. Os resultados obtidos mostraram uma "ótima" concordância entre ambos os métodos. Foi calculada para o método que utiliza CUH uma sensibilidade e especificidade de 98% (93-99%) e 100% (93-100%) respectivamente com uma eficácia de 99%. De acordo com o acima exposto, se conclui que utilizar CUH como substrato para avaliar a presença de EMA é confiável e eficaz para detectar pacientes com doenças celíacas não tratadas.

Ultrastructural characterization of bovine umbilical cord blood cells / Caracterização ultra-estrutural das células sanguíneas do cordão umbilical bovino

The umbilical cord blood (UCB) is an important source of pluripotent stem cells, which motivated researches on ontogeny and transplantation. The morphological characterization of umbilical cord cells is the first step to establish subsequent experiments on these areas. Although some information on humans can be found, no data on UCB is available for bovines. Therefore, this work is the first attempt to conduct an ultrastructural characterization of bovine umbilical cord blood. Blood was collected from the umbilical cord of twenty fetuses by punction of the umbilical vein. Samples were processed for whole leucocytes observation by centrifugation and the buffy coat was collected. Cells were washed and pelleted and prepared according to the standard protocol of the transmission electron microscopy. The presence of cells with morphologic characteristics compatible with the precursors from the erythrocytic, neutrophilic, eosinophilic, basophilic, and lymphocytic lineages was observed. Atypical cells with peculiar morphological features, strongly similar to apoptotic cells, were seen. Bovine neutrophils with three types of cytoplasmic granules were also found in the blood. The ultrastructural characteristics of observed bovine UCB cells where similar to those found in other species, suggesting that bovines could possibly constitute an experimental model for approaches on UCB cells research.

O sangue de cordão umbilical (SCU) é uma importante fonte de células progenitoras pluripotentes, que motiva pesquisas em ontogenia e transplantes. A caracterização morfológica das células de cordão umbilical é o primeiro passo para se estabelecer experimentos subsequentes nessas áreas. Embora algumas informações sobre SCU em humanos possam ser encontradas, não existe nenhuma informação disponível sobre elas em bovinos. Portanto, este trabalho é a primeira tentativa de se conduzir uma caracterização ultra-estrutural do sangue de cordão umbilical bovino. O sangue foi coletado do cordão umbilical de 20 fetos por punção da veia umbilical. As amostras foram processadas para observação dos leucócitos totais por centrifugação pela coleta do botão leucocitário. As células foram lavadas, peletizadas e preparadas de acordo com protocolo padrão para microscopia eletrônica de transmissão. A presença de células com características morfológicas compatíveis com precursores das linhagens eritrocítica, neutrofílica, eosinofílica, basofílica e linfocítica foram observadas. Células atípicas com características morfológicas peculiares muito semelhantes a células em apoptose foram observadas. No sangue do cordão umbilical também foi encontrado neutrófilos bovinos apresentando três tipos de grânulos citoplasmáticos. As características ultraestruturais do SCU bovino foram semelhantes às encontradas em outras espécies, sugerindo que esta espécie possa servir como modelo experimental para abordagens em pesquisa sobre sangue de cordão umbilical.

The mechanism underlying the differentiation of human umbilical cord-derived mesenchymal stem cells into myocardial cells induced by 5-azacytidine.

Objective: To investigate the molecular mechanism underlying the differentiation of human umbilical cord-derived mesenchymal stem cells (hUCMSCs) into myocardial cells induced by 5-azacytidine (5-aza), and to explore the expression and significance of DLL4-Notch signaling in this process. Materials and Methods: hUCMSCs were isolated and purified from the umbilical cords of normal or cesarean term deliveries under sterile conditions. After treatment with 5-aza for 24 h, hUCMSCs was continued to culture, the expression of GATA4 and NKx2.5 at 4 weeks after induction, DLL4 and Notch1 mRNA at 1d, 3d, 5d, 7d after induction were detected. The expression of cardiac troponin I (cTnI) after 4 weeks was determined by immunocytochemistry. Results: hUCMSCs treated with 5-aza were stained positively for cTnI 4 weeks after induction. The expression of Notch1 and DLL4 mRNA in the 5-aza-induced group was stable and significantly higher than that in the control group (mean Ct value for the Notch1 gene: 0.51 ± 0.21 in the 5-aza-induced group vs. 7.85 ± 0.35 in the control group; mean Ct value for the DLL4 gene: 1.60 ± 0.49 in the 5-aza-induced group vs. 12.42 ± 0.73 in the control group). Similar results were observed for Nkx2.5 and GATA4 genes. The expressions of Nkx2.5 and GATA4 mRNA in the 5-aza group were 4.72 ± 0.58 and 3.76 ± 0.06 times higher than that in the control group, respectively, with statistical significance. Conclusions: hUCMSCs can be differentiated into myocardial cells by 5-aza induction in vitro. 5-Aza may affect this process by regulating the expression of GATA4 and Nkx2.5 genes. The DLL4-Notch signal pathway may be involved in this process.

Morphological aspects of buffaloes (Bubalus bubalis) umbilical cord / Aspectos Morfológicos do cordão umbilical de búfalos (Bubalus bubalis)

Buffalo is an important livestock resource, with a great participation in agricultural systems, providing milk, meat, and work power. Umbilical cord is responsible for maternal-fetal nutrients exchange during pregnancy, and its alterations can compromise the fetal development. We investigated ten pregnant uteruses collected from cross-bread buffaloes in different stages of gestation. Pregnancy and fetal age was determined by measuring the apex sacral length and development period was calculated by previously published formula. Umbilical cords were measured for length determination. Umbilical cord vascular net and anastomosis were observed by injection of Neoprene latex. Histological sections of the umbilical cord were studied after stain with HE, picrossirius, toluidine blue, orceine, and PAS reaction. Buffaloes' umbilical cord was formed by two central arteries, an allantois duct and two peripheral veins. The artery wall was composed by large quantity of collagen, elastic fibers, fibroblasts and large number of vasa vasorum. The allantois duct was located between the arteries and presented a great number of small nourishing vessels. Small nourishing vessels should be carefully considered to avoid to be mistaken to the arterials and veins vasa vasorum. Medium length of umbilical cord from buffalos was 11.8cm (minimum of 6.8cm and maximum of 17.4cm).

Búfalo é uma importante fonte de recurso nos rebanhos animais, apresentando uma grande participação na agropecuária, provendo leite, carne e força de trabalho. O Cordão umbilical é responsável pela troca de nutrientes materno-fetais durante a gestação, e suas alterações podem comprometer o desenvolvimento fetal. Nós investigamos dez úteros gravídicos de búfalos de raças cruzadas em fases diferentes de gestação. O período de gestação e a idade fetal foram determinados pelo comprimento ápice sacral, aplicando fórmulas previamente estabelecidas. Posteriormente mediu-se o comprimento do cordão umbilical. A rede vascular do cordão umbilical e anastomoses foram observadas por injeção ou látex de neoprene. O cordão umbilical foi estudado a partir de cortes histológicos, corados por HE, picrossirius, azul de Toluidina, orceína e reação histoquímica de PAS. O cordão umbilical de búfalos é formado por duas artérias centrais, ducto alantóide e duas veias periféricas e apresentam forma de ampulheta. A parede da artéria umbilical é composta por grande quantidade de fibras colágenas e elásticas, fibroblastos e um grande número de vasa vasorum. O ducto alantóide fica alocado entre as artérias e apresenta um grande número de pequenos vasos nutritivos. Os vasos nutritivos devem ser cuidadosamente identificados para evitar-se confundi-los com vasa vasorum. O comprimento médio do cabo de cordão umbilical dos búfalos era 11.8cm (mínimo de 6.8cm e máximo de 17.4cm).

Effects of demineralized bone matrix on proliferation and osteogenic differentiation of mesenchymal stem cells from human umbilical cord.

BACKGROUND: Mesenchymal stem cells or mesenchymal progenitor cells are defined as self-renewable, multipotent progenitor cells with the unlimited capacity to differentiate into multiple lineage-specific cells that form bone, cartilage, fat, and muscle tissues. Demineralized bone matrix (DBM) has been extensively utilized in orthopaedic, periodontal, and maxillofacial applications and widely investigated as a biomaterial to promote new bone formation. OBJECTIVE: To isolate and characterize umbilical cord mesenchymal stem (UCMS) cells and examine the biological activity of DBM in the UCMS cells MATERIAL AND METHOD: UCMS cells were obtained from human umbilical cord culture. Cells were treated with or without DBM over 7 days of culture. Cell proliferation was examined by direct cell counting. Osteogenic differentiation of the UCMS cells was analysed with alkaline phosphatase staining assay. RESULTS: Phenotypic characteristics ofhuman UCMS cells were spindle and stellate shapes with fine homogenous cytoplasm, typically associated with fibroblast-like cells. The control cells (without DBM treatment) exhibited a spindle shape with little extracellular matrix, whereas the DBM treated cells appeared shortened and flattened, and they were surrounded by extracellular matrix. DBM inhibited the growth of the UCMS cells by 50%, as determined by direct cell counting. Morphologic and histochemical studies confirmed that DBM had a strong stimulatory effect on the alkaline phosphatase activities of UCMS cells, a very early marker of cell differentiation into the osteogenic lineage. CONCLUSION: Mesenchymal progenitor cells derived from umbilical cord could differentiate along an osteogenic lineage and thus provide an alternative source for cell-based therapies and tissue engineering strategies.

Hidrops fetal no inmune: presentación de un caso y revisión de la literatura / No immune fetal hydrops: a case report and literature review

El hidrops fetal no inmune (HFNI) es una importante causa de pérdida perinatal, con mortalidad que varía entre 50-100 por ciento. El HFNI es una condición causada por un grupo heterogéneo de patologías. La fisiopatología del desorden que lo produce se conoce en muchos casos. Sin embargo, existen muchos casos en que la causa no se puede detérminar. Presentar un caso de hidrops fetal no inmune, el estudio realizado y la revisión de la literatura. Un caso de hidrops fetal no inmune, fue diagnósticado por ultrasonido antenatal a las 298 semanas de gestación. El feto murió al nacer, el cariotipo de muestra de sangre obtenida del cordón umbilical fue anormal. El examen postmorten fue compatible con Síndrome de Tuner e Higroma Quístico. En el presente caso el HFNI fue causado por la cromosomopatía tipo monosomía X y la anatomía linfática denominada higroma quístico. Todos los casos de HFNI deben ser evaluados prenatalmente para un adecuado diagnóstico y tratamiento cuando la mortalidad es prevenible.

Perfil eletroforético de proteínas séricas do sangue do cordão umbilical de cães / Electrophoretic profile of serum proteins from the umbilical chord blood of dogs

A importância do estudo do sangue do cordão umbilical (SCU) tem sido verificada pela presença de células-tronco hematopoéticas no SCU humano. O modelo experimental em cão tem propiciado informações importantes nos transplantes em humanos. Entretanto, apesar da importância do SCU e do modelo experimental em cães, não existem estudos sobre a eletroforese de proteínas séricas do cordão umbilical canino. No presente protocolo experimental, foi feita a colheita de SCU de 20 neonatos caninos, o qual foi utilizado para o fracionamento eletroforético de suas proteínas. Os valores médios absolutos obtidos para proteínas séricas totais, albumina, alfa, beta e gamaglobulinas no sangue do cordão umbilical de cães ao nascimento foram 3,09±0,59; 1,51±0,38; 0,87±0,17; 0,68±0,12 e 0,03±0,01, respectivamente. Todos os resultados apresentaram-se abaixo dos níveis reportados para animais adultos, devido à passagem das proteínas e suas frações ocorrerem principalmente através do colostro e pela imaturidade hepática. Em todos os traçados eletroforéticos do SCU de cães, foi observada uma pequena curva entre alfaglobulina 2 e betaglobulina 1, não relatada no soro de cães adultos. Portanto, neste experimento, foram observadas diferenças quantitativas no traçado eletroforético das proteínas séricas do SCU de cães, ao nascimento, diferindo-os, neste particular, de cães em diferentes fases da vida pós-colostral.

Morfologia de las celulas en los tumores mixoides / Morphology of the cells in myxoid tumors

Describimos un estudio morfológico de las células en tumores mixoides o con áreas mixoides: fibrosarcomna mixoide, mixoma, liposarcoma mixoide, rabdomiosarcoma embrionario, condroma y condrosarcoma, leiomiosarcoma mixoide, Schwannoma y odontoameloblastoma; y se las compara con los fibroblastos del tejido conectivo reticular del cordón umbilical, del mesénquima embrionario, y del tejido conectivo laxo en procesos inflamatorios. Los métodos utilizados son: H-E, PAS, tricrómico de Masson y la técnica panóptica de Del Río Hortega. Los resultados con la técnica panóptica revelan fibroblastos bipolares con largas prolongaciones. En el mixoma y fibrosarcoma mixoide son estrellados con múltiples prolongaciones y con secreción mucinosa citoplasmática contenida en ampollas o bochas que se deslizan por las prolongaciones hasta resolverse en sus extremos o volcarse al espacio intercelular. En el lipoblastoma mixoide el lipoblasto es también estrellado con múltiples prolongaciones y gotas de lípidos. Los rabdomioblastos son raquetoides con estriaciones transversales; los condroblastos poseen amplio citoplasma con procesos cortos. Los leiomioblastos tienen citoplasma acintado, núcleos en habano y miofibrillas. Las células de Schwann tienen largas y finas prolongaciones opositopolares; los odontoblastos muestran amplio citoplasma con numerosas prolongaciones orientadas hacia la dentina. Estos detalles morfológicos ayudan al diagnóstico diferencial de estos tumores complementados con la inmunocitoquímica