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探讨脐血浆(HUP)能否维持人脐带间充质干细胞(HUCMSCs)的生长。本研究将HUP经盐析、透析等处理后,按10%体积成分培养HUCMSCs,并与同样浓度的胎牛血清(FBS)进行比较,观察细胞的形态,采用流式细胞仪检测细胞表面标志,MTT法检测细胞生长曲线、增殖能力,ELISA检测培养液中bFGF、noggin的分泌浓度。结果发现在HUP和FBS相同培养条件下,HUCMSCs的生长形态、增殖能力均相同,而且在含HUP的培养液中连续培养5代以上仍表现HUCMSCs的表面标志。在HUP和FBS中分泌的bFGF、noggin量分别存在时间-浓度上的差异,且培养超过4h以上的各个时间点HUP组明显高于FBS组。因此我们推测HUP经简单处理后能维持HUCMSCs的长期生长,而且其分泌的细胞因子更适合人胚胎干细胞(hESCs)培养需要,为建立临床级hESCs培养体系作好铺垫。
To investigate whether umbilical cord plasma (HUP) can maintain the growth of human umbilical cord mesenchymal stem cells (HUCMSCs). In this study, HUP was treated with salinization and dialysis. HUCMSCs were cultured in 10% volume fraction and compared with the same concentration of fetal bovine serum (FBS) to observe the cell morphology. Flow cytometry was used to detect cell surface markers, Cell growth curve and proliferation ability were detected by MTT assay. Secretion of bFGF and noggin in culture medium was detected by ELISA. The results showed that under the same culture conditions of HUP and FBS, the growth morphology and proliferation ability of HUCMSCs were the same, and the surface markers of HUCMSCs were still expressed continuously for more than 5 generations in the medium containing HUP. There was a time-concentration difference in the amount of bFGF and noggin secreted in HUP and FBS, respectively. HUP group was significantly higher than that in FBS group at various time points of more than 4 hours. Therefore, we hypothesize that HUP can maintain the long-term growth of HUCMSCs after simple treatment, and that the secreted cytokines are more suitable for the culture of human embryonic stem cells (hESCs) and pave the way for the establishment of a clinical grade hESCs culture system.