Characterization of Chondrogenic Gene Expression and Cartilage Phenotype Differentiation in Human Breast Adipose-Derived Stem Cells Promoted by Ginsenoside Rg1 In Vitro
Characterization of Chondrogenic Gene Expression and Cartilage Phenotype Differentiation in Human Breast Adipose-Derived Stem Cells Promoted by Ginsenoside Rg1 In Vitro
复制标题
体外人参皂苷 Rg1 促进的人乳腺脂肪干细胞中软骨基因表达和软骨表型分化的表征
DOI：
10.1159/000438550
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发表时间：
2015-11
影响因子：
--
通讯作者：
Gang-Yi Chi
中科院分区：
文献类型：
--
作者：
Fang-Tian Xu;Hong-Mian Li;Cheng-Yi Zhao;Zhi-Jie Liang;Min-Hong Huang;Qing Li;Ying-Chao Chen;Gang-Yi Chi
背景/目的：研究和了解人乳腺脂肪干细胞分化为软骨细胞过程中成软骨基因的表达，这是应用该方法进行软骨修复和再生的前提，本研究旨在对人乳腺脂肪干细胞进行鉴定，并在含有人参皂苷Rg1的软骨诱导培养液中检测成软骨基因的表达。方法：根据特异性细胞标记物对第3代人乳腺脂肪干细胞进行鉴定，并对其多向分化能力进行评价。培养的HBASCs分别用基础软骨诱导条件培养液(A组，对照组)或基础软骨诱导培养液+10µg/ml(B组)、50µg/ml(C组)或100µg/.ml人参皂苷Rg1(D组)处理。用CCK-8比色法测定细胞增殖，共9天。诱导2周后，用实时定量聚合酶链式反应(Real-time PCR)检测成软骨基因(II型胶原、XI型胶原、ACP、COMP和弹性蛋白)在各组中的表达。结果：在基础软骨诱导培养液中加入不同浓度的人参皂苷Rg1，在早期(B、C、D组)促进HBASCs的增殖，但导致软骨细胞表型分化，后期其II型胶原(CO-II)、XI型胶原(CO-XI)、酸性磷酸酶(ACP)、软骨寡聚基质蛋白(COMP)和弹性蛋白的表达均高于对照组(A组)。结果显示，人参皂苷Rg1与体外培养的HBASCs的增殖和软骨细胞表型分化存在明显的正相关关系。结论：人乳房脂肪干细胞经第3代培养扩增后仍具有干细胞特性，可作为软骨再生的可行细胞来源。在含有人参皂苷Rg1的条件下进行软骨诱导后，发现HBASCs的软骨形成显著。
Background/Aims: Investigating and understanding chondrogenic gene expression during.the differentiation of human breast adipose-derived stem cells (HBASCs) into chondrogenic.cells is a prerequisite for the application of this approach for cartilage repair and regeneration..In this study, we aim to characterize HBASCs and to examine chondrogenic gene expression.in chondrogenic inductive culture medium containing ginsenoside Rg1. Methods: Human.breast adipose-derived stem cells at passage 3 were evaluated based on specific cell markers.and their multilineage differentiation capacity. Cultured HBASCs were treated either with.basic chondrogenic inductive conditioned medium alone (group A, control) or with basic.chondrogenic inductive medium plus 10 µg/ml (group B), 50 µg/ml (group C), or 100 µg/.ml ginsenoside Rg1 (group D). Cell proliferation was assessed using the CCK-8 assay for a.period of 9 days. Two weeks after induction, the expression of chondrogenic genes (collagen.type II, collagen type XI, ACP, COMP and ELASTIN) was determined using real-time PCR in.all groups. Results: The different concentrations of ginsenoside Rg1 that were added to the.basic chondrogenic inductive culture medium promoted the proliferation of HBASCs at earlier.stages (groups B, C, and D) but resulted in chondrogenic phenotype differentiation and higher.mRNA expression of collagen type II (CO-II), collagen type XI (CO-XI), acid phosphatase (ACP),.cartilage oligomeric matrix protein (COMP) and ELASTIN compared with the control (group.A) at later stages. The results reveal an obvious positive dose-effect relationship between ginsenoside Rg1 and the proliferation and chondrogenic phenotype differentiation of HBASCs.in vitro. Conclusions: Human breast adipose-derived stem cells retain stem cell characteristics.after expansion in culture through passage 3 and serve as a feasible source of cells for.cartilage regeneration in vitro. Chondrogenesis in HBASCs was found to be prominent after.chondrogenic induction in conditions containing ginsenoside Rg1.
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