Differentiation of bone marrow-derived mesenchymal stem cells into chondrocytes using chondrocyte extract
Differentiation of bone marrow-derived mesenchymal stem cells into chondrocytes using chondrocyte extract
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使用软骨细胞提取物将骨髓间充质干细胞分化为软骨细胞
DOI：
10.3892/mmr.2012.996
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发表时间：
2012-10-01
影响因子：
3.4
通讯作者：
Zhang, Qun
中科院分区：
文献类型：
--
作者：
Tang, Xinjie;Sheng, Lingling;Zhang, Qun
使用细胞提取物重编程细胞是用于收获感兴趣的细胞的有效方法。本研究旨在探讨猪骨髓间充质干细胞（BM-MSCs）与软骨细胞提取液共同培养，采用单层和微团培养法，观察BM-MSCs向软骨细胞的分化潜能。通过逆转录-聚合酶链反应（RT-PCR）和免疫荧光法检测软骨特异性标志物。在单层培养中诱导7天后，BM-MSC用链球菌溶血素O（SLO）可逆地透化，SLO是一种能够在哺乳动物细胞质膜中形成大孔的细菌外毒素，并表达软骨细胞特异性基因，如II型胶原（COL II）和聚集蛋白聚糖。COL II蛋白表达呈阳性。而未经SLO处理的BM-MSCs不表达相关基因和蛋白。14天后，重编程的BM-MSCs的转换丢失。通过微团培养，重编程的BM-MSC能够将这种变化维持到第14天。总之，透化的BM-MSC通过与软骨细胞提取物共培养瞬时转分化为软骨细胞。此外，高密度培养方法能够增加维持表型变化的时间。
Reprogramming cells using cell extracts is an effective method for harvesting cells of interest. The aim of this study was to investigate the chondrogenic transdifferentiation potential of swine bone marrow-derived mesenchymal stem cells (BM-MSCs) by culturing with chondrocyte extract, using monolayer and micromass culture. Chondrogenic-specific markers were detected via reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence. After 7 days of induction in monolayer culture, BM-MSCs reversibly permeabilized with streptolysin O (SLO), a bacterial exotoxin that is capable of forming large pores in the plasma membrane of mammalian cells, and expressed chondrocyte‑specific genes such as type II collagen (COL II) and aggrecan. A positive protein expression of COL II was also observed. However, BM-MSCs treated without SLO did not express the related genes and proteins. The transition of reprogrammed BM-MSCs was lost 14 days later. By using micromass culture, reprogrammed BM-MSCs were able to maintain the change until the 14th day. In summary, permeabilized BM-MSCs were transiently transdifferentiated into chondrocytes by co-culturing with the chondrocyte extract. Moreover, a high-density culture method was able to increase the time in which the phenotypic change was maintained.