体外培养骨髓基质细胞增殖和分化的相互关系  

Correlation between proliferation and differentiation of bone marrow stroma cells cultured in vitro

作　　者：刘大鹏[1] 艾合麦提[1] 古丽[2] 穆合甫力[1] 

机构地区：[1]新疆医科大学第一附属医院骨研所,新疆维吾尔自治区乌鲁木齐市830000 [2]新疆医科大学第一附属医院包虫病临床研究所细胞室,新疆维吾尔自治区乌鲁木齐市830000

出　　处：《中国临床康复》2005年第38期10-12,i0001,共4页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:建立一种简单可靠的骨髓基质细胞向成骨细胞转化的体外培养方法,探讨骨髓基质干细胞增殖和分化的相互关系。方法:实验于2003-02/12在新疆医科大学第1附属医院包虫病临床研究所细胞室完成。将兔骨髓基质细胞悬液进行体外培养,传代培养后分组:①普通培养基组传代后仍使用普通培养基培养,每3天换液1次。②条件培养基组,分5种情况,在普通培养基中培养至第3,10,20代后即分别使用条件培养基(Dulbecco’s改良的Eagle’s培养基中加入地塞米松10-7mmol/L,β-甘油磷酸钠10mmol/L,维生素C50mg/L)持续培养30d,每3天换液1次;传代后使用条件培养基持续培养30d,每3天换液1次;传代后使用条件培养基持续培养30d,每4天传代1次。形态学观察采用倒置显微镜;碱性磷酸酶染色采用偶氮偶联法;观察钙结节形成采用Vonkossa’s染色。结果:①各组细胞形态学观察结果:普通培养基组在形态上与条件培养基组细胞无明显差别。②各组细胞钙结节和碱性磷酸酶染色结果:普通培养基培养至第3,10,20代后用条件培养基及传代后用条件培养基(每3天换液)组细胞碱性磷酸酶阳性率达80%以上,表现为成骨细胞形态并形成钙结节,Vonkossa’s染色阳性;普通培养基组及传代后用条件培养基(每4天换液)组细胞的碱性磷酸酶染色为阴性,未能形成钙结节,Vonkossa’s染色阴性。结论:①贴壁筛选法是一种简单可靠的分离骨髓基质细胞的方法。②骨髓基质细胞的分化和增殖是两个对立的状态。AIM： To establish a simple and stable method for the culture of bone marrow stroma cells （BMSc） converting into osteoblast in vitro, and study the mutual relationship between proliferation and differentiation of BMSc. METHODS： The experiment was conducted at the Cell Lab, Clinical Institute of Echinococcosis, First Affiliated Hospital, Xinjiang Medical University between February and December 2003. The cell suspension of rabbits BMSc was cultured in vitro, and after the transfer of culture were divided into groups： ① After transfer of culture, those in the common medium group were cultured in the common medium, changed the liquor once every 3 days. ② Conditional medium group： including 5 kinds, after culturing in the common medium for the 3^rd, 10^th and 20^th generation, the conditional medium （adding 10-7 mmol/L desamethasone, 10 mmol/L β-glycerol phosphate and 50 mg/L vitamin C in DMEM was used for culturing 30 days, changing the liquor once every 3 days. After the transfer of culture, the conditional medium was used to culture continuously for 30 days, changing the liquor once every 3 days. After the transfer of culture, the conditional medium was used to culture continuously for 30 days, transferring once every 4 days. The morphology was observed with invert microscope. The alkaline phosphatase staining was detected with azo-coupling method. The formation of calcium nod was observed with Von kossa＇s staining. RESULTS：① Observation result of cell morphology in every group： There was no obvious difference of the formation between common medium group and conditional medium group. ② Result of calcium nod and alkaline phosphatase staining in every group： After culturing for the 3^rd, 10^th and 20^th in the common medium, the conditional medium was used. The positive rate of alkaline phosphatase .reached over 80% in the conditiona/ medium group （changing the liquor every 3 day） after transferring, manifesting osteoblast formation and forming calcium nod, and the Von kossa＇s sta

关 键 词：骨髓细胞 成骨细胞 组织工程 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]