Abstract

Chorion is the outer fetal membrane around embryo that develops from the trophoblast and the underlying mesenchyme. The objective of this work was to characterize cell lines obtained from chorion of different donors. The expression profile of surface CD markers in three lines derived from three different donors was typical for human mesenchymal stromal cells (MSCs). During long-term cultivation, the cells retained a fibroblast-like morphology. In passages 4–5, the dynamics of the cell cycle was typical of that for normal human cells of mesenchymal origin. Increased cell number in phases of DNA synthesis and mitosis during the logarithmic growth phase subsequently decreased with cell density. In passages 6–7, the pattern of cell cycle distribution altered: cell number in the DNA synthesis phase decreased. Cells accumulated in the G₂/M phase and polyploids number >2n increased. Flow cytometry showed a rapid decrease in the proliferative activity of the cells during their passaging. The population doubling time for one line cell was 40 h in passages 4–5 and increased by passages 6–7 to 52 h. For the other two lines, the doubling time was 80 h in passages 4–5 and the cells died after passages 6–7, ELISA revealed a significant level of VEGF secretion. Karyological analysis showed that chorionic cells in culture have a near-diploid karyotype prone to breakdown of chromosome material. These results show that chorionic MSC are not a reliable object both for use in laboratory studies with lengthy experiments and transplantation for regenerative medicine. However, a high level of VEGF factor secretion makes these cells as a possible source of conditioned medium for therapeutic purposes.

中文翻译：

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从人类绒毛膜建立的间充质基质细胞的表征。

摘要

绒毛膜是从滋养层和潜在的间充质形成的围绕胚胎的胎儿外膜。这项工作的目的是表征从不同供体绒毛膜获得的细胞系。人间充质基质细胞（MSCs）的表面CD标记在来自三个不同供体的三个系中的表达谱是典型的。在长期培养期间，细胞保留了成纤维细胞样的形态。在第4-5代中，细胞周期的动力学是间充质来源的正常人细胞的典型动力学。在对数生长期，DNA合成和有丝分裂阶段的细胞数量增加，随后随着细胞密度的增加而减少。在第6-7代中，细胞周期分布的模式发生了变化：DNA合成阶段的细胞数量减少了。G中积累的细胞₂/ M相和多倍体数> 2n增加。流式细胞仪显示细胞在传代过程中的增殖活性迅速降低。一个传代单元的种群倍增时间在第4-5代为40小时，并在第6-7代至52小时增加。对于其他两行，在第4-5代中倍增时间为80小时，在第6-7代后细胞死亡，ELISA显示VEGF分泌水平显着。核动力分析表明，培养物中的绒毛膜细胞具有易于染色体物质分解的近二倍体核型。这些结果表明绒毛膜MSC既不是可靠的对象，既不能用于进行冗长实验的实验室研究，也不能用于再生医学的移植。然而，