| PART1Comparison of the Biological Characteristics of Human Umbilical Cord and Adipose Tissue Derived Mesenchymal Stem CellsObjective:to investigate the biological characteristics of human umbilical Cord and Adipose Tissue Derived Mesenchymal Stem Cells.Methods:MSCs were isolated from the adipose tissue and umbilical cords from same individual mothers delivering full-term babies. The growth curve of MSCs were drawn according the result of CCK-8test after9successive days of continuous detection. In order to test immunological phenotypes of ASC and UC-MSC, human CD13, CD14, CD44, CD71, CD90, CD105, and CD34, CD45were analyzed using a standard Becton-Dickinson FACSAria instrument and the CellQuest Pro software. The antiapoptotic ability of ASCs and UC-MSCs induced by1×10-6mol/L dexamethasone with48h were analyzed using flowcytery with stained by Annexin-V and PI. In order to acquire the multififferentiation ability of ASC and UC-MSC, differentional induction were performed using the adipogenic, osteogenic and neurogenic induction medium, respectively. At the end of the incubation, adipogenic differentiation was assayed by Oil-Red-O staining for lipid droplets, osteogenic differentiation was assayed by Alizarin red staining for calcium deposition, neurogenic differentiation was assayed by immunofluorescence staining for neural and glial-specific protein expression. The conditional medium were determined by RayBio Biotin Label-based Human Antibody Array I.Results:ASC and UC-MSC were isolated from the related tissue successfully. Morphologies of ASCs and UC-MSCs displayed similar fibroblast-like or spindle-shaped morphology around2days, fused into a sheet, parallel arrangement, and spiral-shaped distribution after6-10days. ASCs and UC-MSCs both exhibited positive surface antigenicity for CD13, CD44, CD71, CD90, and CD105and exhibited negative surface antigenicity for CD14and CD34, CD45. Growth curves of ASCs and UC-MSCs demonstrated that they had the following characteristics in common:in the first12-18hours, cells proliferated slowly and then entered the logarithmic growth phase, which continued for5-6days, and reached cell growth plateau in7-8days, antiapoptotic capacity of ASCs and UC-MSCs were favorable by induced with the high concentration of dexamethasone. ASC and UC-MSC were induced into adipocytes, osteoblast and neuron by the relative induction medium, respectively. The levels of expression of macrophage inflammatory protein2(MIP-2), interleukin6(IL-6),and growth-regulated oncogene (GRO) in UC-MSC-CM were significantly higher than those in ASC-CM, while the levels of expression of CD27and neuregulin in ASC-CM were significantly higher than those of UC-MSC-CM.Conclusion:These results indicate although ASCs and UC-MSCs share considerable similarities in their immunological phenotype and pluripotentiality, certain biological differences do exist, which might have different implications for future cell-based therapy. PART2Conditional supernatant of Human adipose derived Mesenchymal Stem Cells (ASCs) and umbilical cord derived Mesenchymal Stem Cells (UC-MSCs) efficiently induced the apoptosis and differentiation in human glioma cell lineObjective:To investigated the anti-glioma properties of two easily accessible MSCs, namely, human adipose tissue-derived mesenchymal stem cells (ASCs) and umbilical cord-derived mesenchymal stem cells (UC-MSCs).Methods:Proliferation of U251were teat by CCK-8under co-culture by UC-MSC-CM or ASC-CM. The apoptotic of U251induced by ASC-CM or UC-MSC-CM with48h were analyzed using flowcytery with stained by Annexin-V and PI. The expression of apoptosis related genes were analyzed using Quantitative RealTime-PCR. Immunostaining and Cellomics Highcontent Screening (HCS) system assay for glial cell differentiation. Tumor cell infiltration were evaluated by scratch assay and Matrigel invasion assay. Apoptosis related proteins of U251cells were assayed by Raybio AAH-APO-1with induced48h using ASC-CM or UC-MSC-CM.Results:Both type of MSCs can significantly inhibit the growth of human U251cell line, especially for UC-MSCs, which inhibited more than50%growth of U251cells. Annexin and PI flow cytometric assay indicated both type of MSCs can significantly induce apoptosis in human U251cell line. Real-time PCR experiments showed significant upregulation of apoptotic genes of both caspase-3and caspase-9and significant down-regulation of anti-apoptotic genes such as survivin and XIAP after MSC conditioned medium induction. Furthermore, MSCs conditioned medium culture induced rapid and complete differentiation in U251cells. Induced U251cells displayed a relative normal morphology similar to normal glia cells and had significantly decreased tumor infiltration ability.Conclusion:These results indicate human mesenchymal stem cells can efficiently induce both apoptosis and differentiation in U251human glioma cell line. Whereas UC-MSCs are more efficient for apoptosis induction than ASCs, their capability of differentiation induction is not distinguishable from each other. Our findings suggest MSCs themselves have favorable anti-tumor characterics and should be further explored in future glioma therapy. |
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