›› 2018, Vol. 38 ›› Issue (9): 1033-.doi: 10.3969/j.issn.1674-8115.2018.09.006

• Original article (Basic research) • Previous Articles     Next Articles

Comparative cultivation of single cell derived placental mesenchymal stromal cells in vitro

SUN Bao-hangxing1, LU Yi1, WANG Yi-qing1, ZHOU Jia-le1, LIN Meng-lu1, SONG Wei2, JIANG Meng1, BU Jun1   

  1. 1. Department of Cardiology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200240, China; 2. Department of Cardiology, Shuguang Hospital, Shanghai University of Chinese Medicine, Shanghai 200021, China
  • Online:2018-09-28 Published:2018-10-15
  • Supported by:
    National Natural Science Foundation of China, 81270206, 81470391; Shanghai Municipal Education Commission— Gaofeng Clinical Medicine Support, 20172014; Shanghai Jiao Tong University Medical-Engineering Med-X Foundation, YG2014MS49

Abstract: Objective · To explore optimal placental derived mesenchymal stem cells (PMSCs) culture medium using different combination of human platelet lysate (HPL), basic fibroblast growth factor (bFGF) and traditional fetal bovine serum (FBS) for further basic and clinical study. Methods · Single cell derived PMSCs was harvested and incubated with 4 kinds of culture, i.e, FBS, FBS+bFGF, HPL and HPL+bFGF. The morphology, growth state, cell phenotype and multi-energy differentiation were observed. Cells of P1, P2, P3 and P4 generation were counted respectively. The number of units of colony formation was analyzedinoculating P4 cells. Results · It was identified that PMSCs had the biological properties of MSCs. Quantificationally, cell density reached (1.12×107) cells/cm2 in FBS+bFGF group and (1.24×107) cells /cm2 in HPL group (P>0.05) in P4, while those in FBS group and HPL+bFGF group were (5.58×106) cells /cm2 and (8.56×106) cells /cm2, respectively. For cell morphology, the P4 PMSCs of FBS+bFGF and HPL groups kept adherent growth, but the HPL cells could not be adherent in P5 generation. The number of colony was 51/well in FBS+bFGF group, and it was 52/well in HPL group (P>0.05) in P4. The FBS+bFGF group and the HPL group were able to maintain the characteristics of MSCs and the ability of pluripotent differentiation in the P4 generation. Conclusion · PMSCs in P4 cultured in HPL medium can keep the biological characteristics and meet the clinical transplantation requirements in quality and quantity, which are preferable for their low immunogenicity to clinical applications. In long term, PMSCs cultured in FBS+bFGF medium are preferable for the longer lasting characters of MSCs and larger quantity in basic studies. HPL+bFGF medium has no advantage on quality and quantity.

Key words: placental derived mesenchymal stromal cells, cell proliferation, basic fibroblast growth factor, human platelet lysate

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