中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (49): 9147-.doi: 10.3969/j.issn.1673-8225.2010.49.006

• 干细胞培养与分化 • 上一篇    下一篇

不同年龄段大鼠骨髓基质干细胞生物学特性的比较

赵富生1，武庚2，武杨3，金秀东4，张际绯1   

  1. 牡丹江医学院，  1组胚教研室，   2临床技能训练中心，3红旗医院心电图室，4生理教研室，黑龙江省牡丹江市  157011
  • 出版日期:2010-12-03 发布日期:2010-12-03
  • 通讯作者: 张际绯，教授，牡丹江医学院组胚教研室，黑龙江省牡丹江市 157011 zhangjifei777@163.com
  • 作者简介:赵富生★，男，1974年生，黑龙江省人，汉族，2009年佳木斯大学毕业，硕士，讲师，主要从事干细胞与神经损伤修复研究。 zhaofusheng910@163.com
  • 基金资助:

    黑龙江省教育厅科学技术研究项目(11551522)“Neuregulin1转染SCs与BMSCs共移植治疗大鼠糖尿病足及相关机制的研究”。

Comparison of biological characteristics of rat bone marrow stromal stem cells at different ages

Zhao Fu-sheng 1, Wu Geng 2, Wu Yang3, Jin Xiu-dong4, Zhang Ji-fei1   

  1. 1 Department of Histology and Embryology, 2 Clinical Technical Skill Training Center, 4Department of Physiology,Mudanjiang Medical University, Mudanjiang  157011, Heilongjiang Province, China; 3Electrocardiographic Room, Hongqi Hospital, Mudanjiang Medical University, Mudanjiang  157011, Heilongjiang Province, China
  • Online:2010-12-03 Published:2010-12-03
  • Contact: Zhang Ji-fei, Professor, Department of Histology and Embryology, Mudanjiang Medical University, Mudanjiang 157011, Heilongjiang Province, China zhangjifei777@163.com
  • About author:Zhao Fu-sheng★, Master, Lecturer, Department of Histology and Embryology, Mudanjiang Medical University, Mudanjiang 157011, Heilongjiang Province, China zhaofusheng910@163.com
  • Supported by:

    the Science and Technology Research Program of Department of Education of Heilongjiang Province, No. 11551522*

PDF

373

被引次数

14

摘要：

背景：骨髓基质干细胞取材方便，增殖力强，具有多向分化潜能，且可自体移植，是组织工程中理想的种子细胞。年龄等因素对骨髓基质干细胞生物学特性的影响鲜见报道。
目的：比较不同年龄段大鼠骨髓基质干细胞生物学特性的差异，为临床细胞移植提供实验依据。
方法：骨髓基质干细胞供体为SD大鼠。按细胞来源分为3组，即幼年组、成年组和老年组。无菌条件下剪断大鼠股骨两端，用含体积分数为10%胎牛血清的DMEM培养液冲洗骨髓腔，收集骨髓冲洗液，吹打制成单细胞悬液，通过200目的筛网滤过后将其置于培养瓶中培养。取第2代生长均一的细胞，CD44，CD29，CD90 免疫荧光鉴定；观察细胞形态及黏附速度；细胞计数，绘制生长曲线；MTT法检测3组细胞的增殖活力。
结果与结论：CD44，CD29，CD90免疫染色细胞呈阳性；幼年组骨髓基质干细胞的黏附速度快于成年组和老年组；细胞增殖幅度明显 (P < 0.05)；MTT法测定结果显示幼年组骨髓基质干细胞活力强，与另外两组比较差异具有显著性意义(P < 0.05)。提示幼年SD大鼠骨髓基质干细胞增殖快、活力强、可在短时间内大量扩增，是理想的组织工程细胞。

关键词: 年龄, 骨髓基质干细胞, 细胞培养, 细胞增殖, 细胞活力

Abstract:

BACKGROUND: Bone marrow stromal stem cells (BMSCs) have multiple differentiation abilities. BMSCs are easily obtaining, have strong proliferative ability and can do autologous transplantation. Therefore, it is an ideal seed cells in tissue engineering. Few studies have addressed effects of age on biological characteristics of BMSCs.
OBJECTIVE: To compare differences in biological characteristics of BMSCs from rats at different ages, and to provide experimental evidence for clinical cell transplantation.
METHODS: BMSCs were obtained from Sprague-Dawley rats. SD rats were divided into three groups according to the source of cells, namely childhood group, adult group, gerontism group. The femur at both ends was cut under sterile conditions. Bone marrow cavity was washed using DMEM medium containing 10% fetal bovine serum. Bone marrow fluid was collected. Single cell suspension was made, and placed in culture flask after filtration through 200 meshes. The second passage of cells was obtained. CD44, CD29, CD90 protein expression was determined using immunofluorescence staining. Morphology and adhesion rate of BMSCs were observed. The cell growth curve was drawn by cells counting. Activity of BMSCs was detected using MTT assay.
RESULTS AND CONCLUSION: The cells were positive for the CD44, CD29 and CD90 immunostaining. The adhesion rate of childhood group was faster than adult and gerontism groups as well as the proliferation rate (P < 0.05). MTT assay showed that the viability of childhood group was significantly stronger than the other two groups (P < 0.05). Results indicated that BMSCs from the young rats are the ideal tissue engineering cells, for the characters of dynamic and fast amplification in a short time.

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