Morphological characteristics of bone marrow mesenchymal stem cells differentiating into chondrocytes in vitro

doi:10.3969/j.issn.2095-4344.0661

Abstract

Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) can compact to form aggregates during chondrogenic differentiation, but whether the formation of aggregation is a prerequisite for chondrogenic induction is not yet documented.
OBJECTIVE: To observe the morphological characteristics of BMSCs differentiating into chondrocytes.
METHODS: BMSCs were cultured in vitro and passage 6 cells were used for the following experiments. Cells were induced in serum-free LG-DMEM medium as control group or in complete chondrogenic induction medium containing transforming growth factor β3 as induction group for 14 days. Cellular morphology during differentiation process was observed by inverted microscope. Chondrocyte-specific markers, including type II collagen, aggrecan and chondrogenic transcription factor SOX9, were evaluated by immunofluorescence, toluidine blue staining and quantitative RT-PCR.
RESULTS AND CONCLUSION: Compared with the control group, the cells began to lose the typical morphology at day 3 of chondrogenic induction, and then compacted to form mono-layered aggregates and multi-layered aggregates at days 7 and 14, respectively. The aggregates were positive for immunofluorescence and toluidine blue staining, while no staining was detected in the single cells. qRT-PCR results also confirmed the mRNA expression of type II collagen, aggrecan and SOX9 in the cells after chondrogenic induction. Therefore, these abovementioned results suggest that the formation of aggregation in BMSCs can be one of morphological markers of chondrogenic differentiation.

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: Bone Marrow, Mesenchymal Stem Cells, Cell Differentiation, Chondrocytes, Collagen Type II, SOX9 Transcription Factor, Tissue Engineering

CLC Number:

R394.2

Wang Zhi-cong, Liu Yue-hong, Zhou Qing, Chen Xi, Zhou Yu, Sun Hui-jun, Liu Mo-zhen. Morphological characteristics of bone marrow mesenchymal stem cells differentiating into chondrocytes in vitro[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(33): 5281-5285.

Figures/Tables 1

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