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[兔骨髓间充质干细胞的生物学特性及其对不同生长因子的反应]

[Biological characteristics of rabbit bone marrow mesenchymal stem cells and their response to different growth factors].

作者信息

Li Dong, Shen Bai-Jun, Hou Huai-Shui, Shi Qing, Zhang Le-Ling, Ma Xiu-Feng

机构信息

Department of Cryomedicine, Qilu Hospital, Shandong University, Jinan 250012, China.

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2006 Oct;14(5):964-8.

Abstract

This study was aimed to analyze the biological characteristics of rabbit bone marrow mesenchymal stem cells (rBM-MSCs) and their response to different growth factors. Rabbit BM-MSCs were separated from bone marrow mononuclear cells by using adherent cultivation. Biological characteristics were investigated by optical and electron microscopy. Immunophenotype of rBM-MSCs was measured by flow cytometry. The expression of collagen was detected by RT-PCR. Differentiation potential was identified by specific staining and RT-PCR. The response of rBM-MSCs to IL-1, 3, 8 and HGF with different concentrations were tested by MTT. The results showed that the rBM-MSCs gave rise to a population of adherent cells characterized by the presence of a predominant cell type with a typical fibroblast-like morphology and could be cultured for over 15 passages. CD44 was highly expressed on F5 rBM-MSCs (32%) and CD45 was lowly expressed (4.7%). Type I collagen was highly expressed, while type II collagen was lowly expressed and type X collagen was not detected on rBM-MSCs using RT-PCR method. In various conditions inducting differentiation, rBM-MSCs could differentiate into the osteoblast, chondrocyte, adipocyte and neuron-like cells. The rBM-MSCs were sensitive to IL-3, even low concentration (10 ng/ml) of IL-3 could promote the proliferation of rBM-MSCs effectively (>32%, P < 0.01), whereas high concentration IL-3 inhibited it significantly. It is concluded that rabbit BM-MSCs were successfully isolated and culture-expanded. The biological characteristics of rabbit BM-MSCs are similar to those of human and rhesus BM-MSCs. IL-3 with low concentration can promote the proliferation of rBM-MSCs effectively, but high concentration of IL-3 can inhibit their proliferation.

摘要

本研究旨在分析兔骨髓间充质干细胞(rBM-MSCs)的生物学特性及其对不同生长因子的反应。采用贴壁培养法从骨髓单个核细胞中分离兔BM-MSCs。通过光学显微镜和电子显微镜研究其生物学特性。采用流式细胞术检测rBM-MSCs的免疫表型。通过RT-PCR检测胶原蛋白的表达。通过特异性染色和RT-PCR鉴定分化潜能。采用MTT法检测rBM-MSCs对不同浓度的白细胞介素-1、3、8和肝细胞生长因子(HGF)的反应。结果表明,rBM-MSCs产生了一群贴壁细胞,其特征是存在一种主要细胞类型,具有典型的成纤维细胞样形态,并且可以传代培养超过15代。CD44在F5 rBM-MSCs上高表达(32%),而CD45低表达(4.7%)。使用RT-PCR方法检测发现,rBM-MSCs上I型胶原蛋白高表达,而II型胶原蛋白低表达,未检测到X型胶原蛋白。在各种诱导分化条件下,rBM-MSCs可分化为成骨细胞、软骨细胞、脂肪细胞和神经元样细胞。rBM-MSCs对IL-3敏感,即使低浓度(10 ng/ml)的IL-3也能有效促进rBM-MSCs的增殖(>32%,P<0.01),而高浓度的IL-3则显著抑制其增殖。结论是兔BM-MSCs成功分离并培养扩增。兔BM-MSCs的生物学特性与人和恒河猴BM-MSCs相似。低浓度的IL-3能有效促进rBM-MSCs的增殖,但高浓度的IL-3会抑制其增殖。

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