In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells
In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells
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DOI：
10.1006/excr.1997.3858
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发表时间：
1998-01-10
影响因子：
3.7
通讯作者：
Yoo, JU
中科院分区：
文献类型：
--
作者：
Johnstone, B;Hering, TM;Yoo, JU
本研究建立了一种促进兔骨髓间充质祖细胞向软骨细胞分化的培养体系。首先通过单层培养分离骨髓抽吸物中获得的细胞，然后转移至试管中，并使其在化学成分确定的培养基中形成三维聚集体。在培养基中加入10（-7）M地塞米松诱导聚集体内细胞的软骨形成分化，如甲苯胺蓝异染的出现和早在开始三维培养后7天的II型胶原的免疫组织化学检测所证明的。21天后，整个聚集体的基质含有II型胶原。培养14天，也有证据表明X型胶原蛋白存在于基质中，细胞形态学上类似于肥大软骨细胞。然而，仅在所用的骨髓细胞制剂的约25%中实现了软骨形成分化。相反，加入转化生长因子-β 1（TGF-β 1）后，无论是否存在10（-7）M地塞米松，所有骨髓细胞制剂均诱导软骨形成。RT-PCR实验结果表明，在聚集后7天，IIA和IIB型胶原mRNA都被检测到，X型胶原mRNA也被检测到。相反，I型胶原mRNA的表达在预聚集细胞中检测到，但在聚集后7天不再检测到。这些结果为成人哺乳动物骨髓来源的祖细胞体外软骨分化提供了组织学、免疫组化和分子证据。(C)北京：科学出版社.
A culture system that facilitates the chondrogenic differentiation of rabbit bone marrow-derived mesenchymal progenitor cells has been developed. Cells obtained in bone marrow aspirates were first isolated by monolayer culture and then transferred into tubes and allowed to form three-dimensional aggregates in a chemically defined medium. The inclusion of 10(-7) M dexamethasone in the medium induced chondrogenic differentiation of cells within the aggregate as evidenced by the appearance of toluidine blue metachromasia and the immunohistochemical detection of type II collagen as early as 7 days after beginning three-dimensional culture. After 21 days, the matrix of the entire aggregate contained type II collagen. By 14 days of culture, there was also evidence for type X collagen present in the matrix and the cells morphologically resembled hypertrophic chondrocytes. However, chondrogenic differentiation was achieved in only approximately 25% of the marrow cell preparations used. In contrast, with the addition of transforming growth factor-beta 1 (TGF-beta 1), chondrogenesis was induced in all marrow cell preparations, with or without the presence of 10(-7) M dexamethasone. The induction of chondrogenesis was accompanied by an increase in the alkaline phosphatase activity of the aggregated cells, The results of RT-PCR experiments indicated that both type IIA and IIB collagen mRNAs were detected by 7 days postaggregation as was mRNA for type X collagen. Conversely, the expression of the type I collagen mRNA was detected in the preaggregate cells but was no longer detectable at 7 days after aggregation, These results provide histological, immunohistochemical, and molecular evidence for the in vitro chondrogenic differentiation of adult mammalian progenitor cells derived from bone marrow. (C) 1998 Academic Press.