白血病抑制因子联合bFGF对人BMSCs增殖及分化的影响

发布时间：2018-10-24 07:12

【摘要】：目的研究白血病抑制因子(leukemia inhibitory factor,LIF)联合bFGF对人BMSCs(human BMSCs,hBMSCs)增殖及分化的影响。方法取第4代hBMSCs根据培养条件不同分为4组:A组为对照组,加入完全培养基(含10%FBS的α-MEM)常规培养;B组加入含10 ng/mL LIF的完全培养基;C组加入含10 ng/mL bFGF的完全培养基;D组加入含10 ng/mL LIF和10 ng/mL bFGF的完全培养基。采用细胞计数试剂盒8(cell counting kit 8,CCK-8)测定各组第4代hBMSCs生长曲线;倒置相差显微镜观察各组hBMSCs形态变化;流式细胞术检测各组第8代hBMSCs表面标志CD44、CD90、CD19、CD34表达情况。结果 4组细胞生长曲线均近似S形,且细胞增殖速率表现为D组C组B组A组。各组细胞形态观察发现A组细胞较早出现衰老和分化现象;B组细胞早期形态维持较好,但增殖相对缓慢,后期出现部分衰老细胞;C组少量细胞呈神经样细胞分化,但增殖较快;D组细胞增殖迅速,且能长期维持hBMSCs形态特征。流式细胞术检测示,A、C组骨髓基质标志CD44、CD90表达较B、D组低;各组造血细胞标志CD19、CD34均呈阴性,组间差异不明显。结论 LIF联合bFGF不仅可以维持hBMSCs的多向分化潜能,而且能够促进其快速增殖。
[Abstract]:Objective To study the effect of bFGF on proliferation and differentiation of human BMSCs and hBMSCs. Methods The 4th generation hBMSCs were divided into 4 groups according to the culture conditions: group A was control group, complete culture medium (containing 10% FBS) routine culture was added, B group was added with 10 ng/ mL LIF complete medium, and group C was added with 10 ng/ mL bFGF complete medium. A complete medium containing 10 ng/ mL LIF and 10 ng/ mL bFGF was added to the D group. Cell counter kit 8 (CCK-8) was used to determine the growth curve of hBMSCs in each group; the morphological changes of hBMSCs were observed by reversed phase contrast microscope; and the expression of CD44, CD90, CD19 and CD34 on the 8th generation hBMSCs was detected by flow cytometry. Results The growth curves of the four groups were approximately S-shaped, and the cell proliferation rate was expressed as group B (group A). The cell morphology of group B showed that group A had earlier aging and differentiation; the early morphology of group B was good, but the proliferation was relatively slow, and some aging cells appeared later; small number of cells in group C differentiated into neural-like cells, but the proliferation was faster; the proliferation of group D cells was rapid. and the morphological characteristics of hBMSCs can be maintained for a long time. The expression of CD44 and CD90 in bone marrow stromal cells of group A and C was lower than that in group D, and CD19 and CD34 were negative in each group, and the difference between groups was not obvious. Conclusion LIF combined with bFGF can not only maintain the multipotent differentiation potential of hBMSCs, but also can promote the rapid proliferation of hBMSCs.
【作者单位】：九江学院基础医学院;南昌大学研究生院医学部;南昌大学第二附属医院骨科;
【基金】：国家自然科学基金资助项目(81260283) 江西省自然科学基金资助项目(20122BAB205043) 江西省教育厅科学技术研究项目(赣教技字10737号)~~
【分类号】：R329.2

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