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BACKGROUND:Classic media and fetal bovine serum are commonly used in the culture of umbilical cord mesenchymal stem cells, but the potential risk of serum culture limits its clinical application. OBJECTIVE:To use human platelet lysate alternative to fetal bovine serum for large-scale production of mesenchymal stem cells for therapeutic applications. METHODS:Human platelet lysate was prepared by repeated freezing and thawing, centrifugation, filtration, and concentration. Human umbilical cord mesenchymal stem cells were cultured in Iscove’s Modified Dulbecco’s Medium containing 5%concentrated platelet lysate (experimental group) or 10%fetal bovine serum (control group). After separation and enzymatic digestion, human umbilical cord mesenchymal stem cells were subcultured at a concentration of 3 000/cm2 up to the fifth generation. Then, cellmorphology and diameter, immune phenotype, osteogenic and adipogenic differentiation, and cloning efficiency were detected and compared between two groups. RESULTS AND CONCLUSION:Human umbilical cord mesenchymal stem cells cultivated in human platelet lysate-supplemented media showed a smal er size and more elongated morphology than those in fetal bovine serum-supplemented media. Colony forming unit-fibroblast analyses further showed no significant differences in colony efficiency. Human umbilical cord mesenchymal stem cells cultivated in human platelet lysate-supplemented media showed an increase of proliferation capacity;whereas, similar immunophenotypes remained in the two groups. In vitro assays revealed intact differentiation potential. Moreover, human umbilical cord mesenchymal stem cells cultivated in human platelet lysate-supplemented media showed a significantly higher capacity to differentiate towards osteocytes, indicating human platelet lysate is an alternative to fetal bovine serum for low-density production of mesenchymal stem cells for therapeutic applications.
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BACKGROUND:Preliminary experiments have reported the influence of serum and nerve growth factor on olfactory ensheathing cells proliferation in vitro, but there are less studies concerning choice of serum concentration and growth time for in vitro culture of olfactory ensheathing cells. OBJECTIVE:To find out the influence of different blood serum concentration and growth time on olfactory ensheathing cells from the olfactory mucosa of adult rats based on the growth curve of olfactory ensheathing cells. METHODS:The olfactory ensheathing cells from the olfactory mucosa of adult rats were separated, culture and identified in vitro. Sulforhodamine B and microplate reader were employed to measure absorbance values and plot growth curve of olfactory ensheathing cells. RESULTS AND CONCLUSION:When cultured for the same time in blood serum of different concentrations, absorbance values, especial y in the groups 10%, 20%, 30%, 40%, tended to increase with time except the 0%group. When cultured in the same serum for different time, absorbance values increased within the first 9 days, then promoted rapidly in the groups 10%, 20%, 30%, 40%at 13 days, entered the plateau phase at 19 days, and decreased at 23 days;meanwhile, in the other groups (50%, 60%, 70%, 80%, 90%) the absorbance values peaked at the 13th day and then decreased gradual y. These findings indicate that different serum concentrations and different growth time in vitro affect cellgrowth and survival of olfactory ensheathing cells significantly, which should be ful y considered when cells are cultured in an in vitro condition.
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BACKGROUND:At present, heterologous serum as a medium is a common method for culture of bone marrow mesenchymal stem cells, but it is limited by the disease transmission between species, the potential immune rejection and even controversial ethic issues. This method is also contrary to the requirements of the Ministry of Health. Autologous platelet-rich plasma is a kind of whole blood extract, containing a variety of growth factors. OBJECTIVE:To explore the osteogenic differentiation of rabbit bone marrow mesenchymal stem cells cultured in autologous platelet rich plasma alternative to traditional heterogeneous serum-free culture. METHODS:8 mL bone marrow from the rabbit iliac crest was extracted and anti-coagulated with heparin, and then bone marrow mesenchymal stem cells were isolated using density gradient centrifugation. The cells were divided into autologous platelet rich plasma group (10%autologous platelet rich plasma) and fetal bovine serum group (10%fetal bovine serum). At the passage 4, the cells in the two groups were respectively subdivided into experimental and control groups. Experimental groups were subjected to osteogenic induction, while no change was done in the control groups. cellproliferation was determined by using growth curves;the activity of alkaline phosphatase was detected to adjust the osteogenic differentiation of cells in different groups. RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells at different generations al showed good proliferation. At 12 days of autologous platelet rich plasma culture, the activity of alkaline phosphatase in the passage 4 cells in the experimental group was significantly higher than that in the control group;while the activity of alkaline phosphatase in the autologous platelet rich plasma group was significantly higher than that in the fetal bovine serum group (P<0.01). These findings indicate that autologous platelet rich plasma as a substitute of xenogeneic serum for culture of bone marrow mesenchymal stem cells is a method characterized as safe and reliable, simple operation, high-purity active induction.
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Objective To observe the clinical curative effect and security of the deproteinised calf serum enteric capsules on nerve functions defect caused by cerebral isehemia damage.Methods A randomized,double-blind,multicentric trial design was conducted.Treatment group:based on routine treatment,the patients were given deproteinised calf serum enteric capsules for 90 days.Control group: based on routine treatment,the patients were given placebo for 90 days.The NIHSS,BI,mRS measuring scale were used for evaluation on the 7th,14th and 28th day before and after treatment.BI and mRS measuring scale were used for evaluation on the 90th day.The change of numerical value was inspected before and after treatment.Results Before treatment,there was no significant difference in the NIHSS,BI and mRS measuring scale evaluation between the groups of treatment and control.After treatment,there were significant differences in the NIHSS,BI and mRS measuring scale evaluation between the groups of treatment and control on the 14th and 28th day (P