[1]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):408-411,418.
 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,(04):408-411,418.
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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()
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《南方医科大学学报》[ISSN:/CN:]

卷:
期数:
2004年04期
页码:
408-411,418
栏目:
出版日期:
2004-04-01

文章信息/Info

Title:
Induced differentiation of adult human bone marrow derived mesenchymal stem cells in vitro toward osteoblasts
作者:
刘晓静1 任高宏2 廖华1 余磊1 原林1
1. 第一军医大学解剖教研室, 广东广州, 510515;
2. 第一军医大学南方医院全军创伤骨科中心, 广东广州, 510515
Author(s):
LIU Xiao-jing1 REN Gao-hong2 LIAO Hua1 YU Lei1 YUAN Lin1
1. Department of Anatomy, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China;
2. Center of Orthopaedics and Traumatology of PLA, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China
关键词:
成骨细胞细胞培养间充质干细胞骨组织工程
Keywords:
osteoblastscell culturemesenchymal stem cellsbone tissue engineering
分类号:
Q254;Q421
摘要:
目的 研究体外培养的成人骨髓间充质干细胞(MSC)在特定的条件下定向诱导分化为成骨细胞,探讨其作为骨组织工程种子细胞的可行性和应用价值。方法 抽取健康成年志愿者骨髓,在含10%胎牛血清的高糖DMEM培养基中采用全骨髓培养法,培养传代后改用含地塞米松(1×10-8 mol/L)、β-甘油磷酸钠(10 mmol/L)和维生素C(50 mg/L)的条件培养基培养,在相差显微镜和电镜下观察细胞形态,免疫组化检测Ⅰ型胶原,Gomori钙钴法进行碱性磷酸酶(AP)染色、von Kossa法进行钙结节染色,同时测定细胞内AP(碱性磷酸酶)含量变化,并进行统计学分析。结果 原代和传代培养的细胞具有活跃的增殖能力,诱导培养2~3周后,透射电镜下观察见大量扩张的粗面内质网、高尔基体和线粒体,细胞核较为幼稚。Ⅰ型胶原染色、AP及钙结节染色等均为强阳性;AP活性明显增强(P<0.05)。结论 MSC取材安全方便,易于诱导分化为成骨细胞,有望成为理想的骨组织工程种子细胞来源,本实验方法可作为骨组织工程种子细胞培养的常规方法之一。
Abstract:
Objective To investigate the feasibility of inducing in vitro mesenchymal stem cells (MSCs) derived from adult human bone marrow differentiate into osteoblasts and potential applicability of the MSCs as the seed cells in tissue engineering. Methods Adult human bone marrow was collected from the healthy adult volunteers to obtain the MSCs, which, after in vitro culture in DMEM supplemented with 10% fetal bovine serum and incubation under standard condition, were induced to differentiate into osteoblasts in DMEM containing dexamethasone (1×10-8mol/L), beta-sodium glycerophosphate (10 mmol/L) and ascorbic acid (50 mg/L). Proliferation and differentiation of the MSCs were observed continually under inverted phase-contrast microscope and transmission electron microscope. The collagen typeⅠwas detected by immunohi- stochemistry, alkaline phosphatase (AP) in the MSCs stained by Gomori, the calcified nodules were stained by von Kossa method, and the changes in the content of AP were measured. Results The MSCs proliferated rapidly in in vitro culture and after a 2- to 3-week induction, the cells began to generate large amount of enlarged endoplasmic reticulum, Golgi complexes and mitochondria, with immature cell nuclei. Positive staining for collagen typeⅠand strong reaction for AP and calcified nodules were observed. Increasing AP secretion by the MSCs was seen as the time of induction prolonged (P<0.01). Conclusions Human bone marrow-derived MSCs can be induced to differentiate into osteoblasts through relatively simple procedures, which provide ideal autogenous source of seed cells for bone tissue engineering. The method adopted in this experiment may be used for routine culture of the seed cells for bone tissue engineering.

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备注/Memo

备注/Memo:
收稿日期:2003-11-9。
基金项目:广东省攻关课题(2001A302020401)
作者简介:刘晓静(1975-),女,医学硕士,2003年毕业于第一军医大学,研究方向:组织工程,电话:020-61648198,E-mail:Liuxj@fimmu.com
更新日期/Last Update: 1900-01-01