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首页> 外文期刊>Cell biology international. >Induction of chondrogenic differentiation of mouse embryonic mesenchymal stem cells through an in vitro pellet model
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Induction of chondrogenic differentiation of mouse embryonic mesenchymal stem cells through an in vitro pellet model

机译：通过体外沉淀模型诱导小鼠胚胎间充质干细胞的软骨分化

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This study employed transforming growth factor beta 3 (TGF-3) induction of the C3H10T1/2 mesenchymal stem cell (MSC) line to construct an in vitro chondrogenic differentiation model for MSCs. A C3H10T1/2 MSC cell line was cultured, amplified, and the seventh generation of cells was centrifuged to construct pellets, which were divided into a non-induced group and an induced group (treated with TGF-3, vitamin C, dexamethasone, and ITS). Specimens were taken after 7, 14, 21, and 28 days under non-induced and induced culture to compare these two groups by Alcian Blue staining, collagen type II immunohistochemical staining, and transmission election microscopy (TEM) on days 21 and 42. Cell pellets in the non-induced group were smaller than those in the induced group on days 7, 14, 21, and 28 with the pellet morphology of the induced group being more regular and nearly spherical. Alcian blue staining in the induced group was consistently stronger than that in non-induced group across all time points, and type II collagen immunohistochemical IOD values were significantly higher in the induced group over the non-induced group across all time points. On days 21 and 42, TEM revealed that the induced group displayed greater karyokinesis and a higher euchromatin ratio compared to the non-induced group. This specially constructed pellet model treated with TGF-3-containing chondrogenic medium can effectively promote chondrogenic differentiation of C3H10T1/2 MSC cells in vitro. This in vitro pellet model should be of value in providing a preliminary cell model reference for further studies of the mechanism of chondroblast differentiation of stem cells.
机译：这项研究采用了C3H10T1 / 2间充质干细胞（MSC）系的转化生长因子β3（TGF-3）诱导，以构建MSC的体外软骨形成分化模型。培养，扩增C3H10T1 / 2 MSC细胞系，并将第七代细胞离心以构建沉淀，将其分为未诱导组和诱导组（分别用TGF-3，维生素C，地塞米松和它的）。在未诱导和诱导培养的第7、14、21和28天后采集标本，以通过Alcian Blue染色，II型胶原免疫组化染色以及第21和42天的透射电子显微镜（TEM）比较这两组。在第7、14、21和28天，未诱导组的小丸比诱导组小，诱导组的小丸形态更规则且接近球形。在所有时间点上，诱导组的阿尔辛蓝染色始终强于未诱导组，并且在所有时间点，诱导组的II型胶原免疫组织化学IOD值均显着高于未诱导组。在第21天和第42天，TEM显示，与未诱导组相比，诱导组表现出更大的核运动能力和较高的常染色质比率。这种特殊构建的颗粒模型用含TGF-3的软骨形成培养基处理可以有效地促进C3H10T1 / 2 MSC细胞的软骨分化。这种体外沉淀模型对于为进一步研究干细胞软骨母细胞分化的机制提供初步的细胞模型参考具有价值。

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