[1]祝旭龙,颜谭,姚维杰,等.大鼠骨髓间充质干细胞的分离与培养方法优化[J].南方医科大学学报,2014,(11):1621.
点击复制

大鼠骨髓间充质干细胞的分离与培养方法优化()
分享到:

《南方医科大学学报》[ISSN:/CN:]

卷:
期数:
2014年11期
页码:
1621
栏目:
出版日期:
2014-11-15

文章信息/Info

Title:
Optimization of the method for isolating and culturing rat mesenchymal stem cells
作者:
祝旭龙颜谭姚维杰王永恒程冲向俊西吕毅高庆东李建辉
关键词:
骨髓间充质干细胞分离与培养肝细胞胆管上皮细胞大鼠
Keywords:
bone marrow mesenchymal stem cell isolation and culture hepatocytes biliry epithelial cells
摘要:
目的建立一个分离、培养、纯化大鼠的骨髓间充质干细胞的方法。方法使用贴壁法分离大鼠的骨髓间充质干细胞,通过
培养初期的频繁换液和传代过程中减少胰蛋白酶的暴露时间,达到骨髓间充质干细胞的纯化。流式细胞仪检测细胞表面标志
物,MTT法检测细胞生长状况,暴露于不同的诱导培养基使细胞成骨、成脂,并向肝胆系分化。结果分离所得的细胞阳性表达
CD29、CD44、CD90,而对于造血细胞标志物CD45呈阴性。在成骨、成脂培养基的诱导下,细胞ALP、茜素红、油红O染色均呈
阳性,并发现传至10代的细胞仍保持分化功能。将骨髓间充质干细胞贯续暴露于不同的细胞因子和激素,得到了表达肝细胞、
胆管上皮细胞特异标志的子代细胞。结论通过改良后的方法可以高效、简洁地分离出形态均一的大鼠骨髓间充质干细胞,并
能够向多胚层的细胞分化。
Abstract:
Objective To optimize the protocols for isolation and culture of mesenchymal stem cells from rat bone marrow
(BMSCs). Methods BMSCs were isolated by adherence to plastic with frequent medium change and reduced trypsinization
time. The cell growth curves were drawn and the surface markers of BMSCs were detected by flow cytometry. The cells were
induced to differentiate into osteogenic, adipogenic, hepatic and cholic lineages. Results The cells isolated using this method
were positive for CD29, CD44, and CD90 and negative for the hematopoietic surface markers CD45. The osteogenic and
adipogenic differentiation of the BMSCs was verified by alkaline phosphatase staining, Alizarin red staining and Oil red
staining. The cell subcultures up to passage 10 maintained capacities of differentiation into osteogenic and adipogenic lineages.
The BMSCs induced with sequential addition of growth factors, cytokines and hormones differentiated into cells expressing
hepatocyte- and cholangiocyte-specific markers. Conclusion The optimized method allows efficient isolation of homogenous
populations of MSCs from rat bone marrow, which can be induced into multiple cell lineages.

相似文献/References:

[1]张俊,谢珊珊,韩晓霞,等.血管内皮生长因子对骨髓源间充质干细胞增殖的影响及其信号机制[J].南方医科大学学报,2011,(10):1697.
[2]顾春瑜,王前,郑磊,等.小鼠骨髓耐受性树突状细胞的体外培养与鉴定[J].南方医科大学学报,2005,(08):959.
 GU Chun-yu,WANG Qian,ZHENG Lei,et al.In vitro culture and identification of tolerogenic dendritic cells from mouse bone marrow[J].,2005,(11):959.
[3]郑莉,周美娟,丁振华.湿热环境对辐射损伤动物骨髓和脾脏的影响[J].南方医科大学学报,2004,(12):1422.
 ZHENG Li,ZHOU Mei-juan,DING Zhen-hua.Effect of hot and humid environment on bone marrow and spleen of mice exposed to γ irradiation[J].,2004,(11):1422.
[4]刘晓静,任高宏,廖华,等.成人骨髓间充质干细胞向成骨细胞定向诱导分化的实验研究[J].南方医科大学学报,2004,(04):408.
 LIU Xiao-jing,REN Gao-hong,LIAO Hua,et al.Induced differentiation of adult human bone marrow derived mesenchymal stem cells in vitro toward osteoblasts[J].,2004,(11):408.
[5]柳太云,张成,肖露露,等.人脐血单个核细胞诱导分化为神经元样细胞[J].南方医科大学学报,2005,(02):152.
 LIU Tai-yun,ZHANG Cheng,XIAO Lu-lu,et al.Induced differentiation of human cord blood monocytes into neuron-like cells in vitro[J].,2005,(11):152.
[6]柳太云,李经伦,姚晓黎,等.3H-TdR标记人骨髓间充质干细胞移植治疗mdx鼠的实验研究[J].南方医科大学学报,2005,(05):498.
 LIU Tai-yun,LI Jing-lun,YAO Xiao-li,et al.Transplantation of 3H-thymidine-labeled human bone marrow-derived mesenchymal stem cells in mdx mice[J].,2005,(11):498.
[7]赵文秀,张磊,刘建明,等.小鼠脂肪间充质干细胞的分离培养及其在肝内归巢[J].南方医科大学学报,2013,(08):1151.
[8]廖军义,周年,林良波,等.联合表达BMP2和Sox9促进体外培养间充质干细胞成软骨分化[J].南方医科大学学报,2014,(03):317.
[9]袁茵,陈丹亮,陈璇,等.人脐带间充质干细胞通过激活Notch信号通路促进白血病细胞株Jurkat的体外存活[J].南方医科大学学报,2014,(04):441.
[10]聂瑛洁,罗利梅,查艳,等.系统性红斑狼疮患者血浆降低间充质干细胞对B淋巴细胞的抑制作用[J].南方医科大学学报,2016,(08):1090.

更新日期/Last Update: 1900-01-01