Biological characteristics of three kinds of human placenta-derived mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.1600

Abstract

Abstract:

BACKGROUND: The placental tissue structure is complex, including the amniotic membrane, chorion, and decidua from the mother. Mesenchymal stem cells derived from different tissues of the same placenta have been reported to have similar biological characteristics. To date, there is no study regarding quantitative comparison of differentiation potential and immunomodulatory function of mesenchymal stem cells derived from different tissues of human placenta.
OBJECTIVE: To investigate the biological characteristics including differentiation potential and immunomodulatory function of mesenchymal stem cells derived from different tissues of human placenta.
METHODS: The amnion-, chorion-, and decidua-derived mesenchymal stem cells were isolated from the placental tissue of a baby boy by enzymatic digestion method. The biological characteristics of these three kinds of mesenchymal stem cells were systematically investigated including cell morphology, immunophenotypes, karyotypeanalysis, adipogenic and osteogenic differentiation potential, and Treg cells proliferation capacity.
RESULTS AND CONCLUSION: All three kinds of mesenchymal stem cells showed fibroblast-like morphology and expressed the surface markers of mesenchymal stem cells with high expressions of CD73, CD90 and CD105, as well as low expressions of CD14, CD19, CD34, CD45 and HLA-DR. The karyotypes of the amnion- and chorion-derived mesenchymal stem cells were the same as the fetus, and decidua-derived mesenchymal stem cells had the same karyotype as the mother. There were significant differences in adipogenic differentiation capacity between three kinds of mesenchymal stem cells (amnion-derived mesenchymal stem cells > chorion-derived mesenchymal stem cells > decidua-derived mesenchymal stem cells; P < 0.05). In contrast, the osteogenic differentiation capacity of amnion-derived mesenchymal stem cells was remarkably higher than that of decidua-derived mesenchymal stem cells (P < 0.05). The amnion- and chorion-derived mesenchymal stem cells had the higher potential of Treg cell proliferation induction than decidua-derived mesenchymal stem cells. These findings indicate that three sources of human placenta-derived mesenchymal stem cells have different karyotypes, adipogenic and osteogenic differentiation potential, and immunomodulatory capability, providing a variety of ideal seed cell sources for disease treatment.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Placenta, Mesenchymal Stem Cells, Amnion, Chorion, Decidua, Immunomodulation, Cell Differentiation, Tissue Engineering

CLC Number:

R459.9

Wang Liudi, Liu Wei, Xie Yuanyuan, Gao Tianyun, Huang Feifei, Wang Bin. Biological characteristics of three kinds of human placenta-derived mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(9): 1377-1383.

Figures/Tables 1

2.1 不同胎盘组织来源间充质干细胞的形态接种48 h后，大部分细胞贴壁，72 h换液时可见少量细胞集落。细胞呈梭形或多角形，3-5 d后增殖加快，呈平行或漩涡状排列。培养5-7 d细胞可铺满瓶底80%以上。多次传代后细胞形态较为均一，均呈现成纤维形态，符合间充质干细胞形态特点，见图1。 2.2 不同胎盘组织来源间充质干细胞的表面标志物流式细胞仪分析结果显示，羊膜、绒毛膜以及蜕膜来源细胞均高表达CD73、CD90和CD105，大于95%；低表达CD14、CD19、CD34、CD45和HLA-DR，小于2%，与间充质干细胞表面标志物通用标准一致，见图2。 2.3 不同胎盘组织来源间充质干细胞的核型分析 G-带染色结果显示3种细胞染色体结构正常。羊膜、绒毛膜来源间充质干细胞的核型为46，XY，表明细胞来源于男性胎儿组织；蜕膜来源间充质干细胞的核型为46，XX，表明细胞来源于母体组织，见图3。 2.4 不同人胎盘组织来源间充质干细胞的成脂、成骨分化能力比较成脂诱导培养21 d后，油红O染色可见红色的脂滴，见图4；成骨诱导培养21 d后，茜素红染色均出现深红色的钙盐结节，见图5。统计分析结果表明：3种细胞都具有成脂成骨分化潜能，其中蜕膜来源间充质干细胞的成脂分化能力最强，依次为蜕膜来源间充质干细胞>绒毛膜来源间充质干细胞>羊膜来源间充质干细胞，各组间差异有显著性意义；而这3种细胞的成骨分化能力与成脂分化相反，成骨分化能力依次为羊膜来源间充质干细胞>绒毛膜来源间充质干细胞>蜕膜来源间充质干细胞，但仅羊膜来源间充质干细胞与蜕膜来源间充质干细胞的成骨分化能力差异有显著性意义(P < 0.05)。 2.5 不同人胎盘组织来源间充质干细胞对调节性T细胞增殖能力的影响 3种间充质干细胞与外周血单个核细胞在重组人白细胞介素2刺激下体外共培养5 d，均促进CD4+CD25+Foxp3+调节性T细胞增殖，见图6。羊膜、绒毛膜来源间充质干细胞高于蜕膜来源间充质干细胞，差异有显著性意义。绒毛膜来源间充质干细胞稍高于羊膜来源间充质干细胞，但是差异无显著性意义。"

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