小型猪自体骨髓间充质干细胞移植治疗急性心肌梗死

• 作者: 王士雯 程瑞
• 来源期刊: 世界急危重病医学杂志
• 学科: 医学
• 关键词: 急性心肌梗死 细胞移植治疗 骨髓间充质 lymphocyte biological different methodology cells 小型猪 augmented mol/L 5-AZA TROPONIN MSCs protein 自体 DESMIN DESMIN little effect and The the fetal serum group
• 年，卷（期）: 2005,（4）
• 页码范围: 800-804
• 页数: 5页
• 分类号: R542.22

摘要

Objective To establish the method for isolation, culture and proliferation of swine bone marrow MSCs and investigate the potential and contribution of cell proliferation MSCs derived from swine bone marrow to cardiomyogenic differentiation after exposure to different concentration of 5-azacytidine (5-aza) in vitro. Methods 10ml bone marrow was taken from swine,and mononuclear cells (MNCs) were freshly isolated by 1.077 g/ml Ficoll lymphocyte separating medium using density-gradient centrifugation. The 2nd generated MSCs were exposured in medium mixed with different concentration of 5-aza (3,5,10 proM/L) for 24 h cellular differentiation ,than cultured in complete medium for 4 weeks together with control group. Desmin and cardiac-specific cardiac troponin I (cTnI) were examined respectively with immunohischemistry at 28 days and the uhramicrostructure of induced cells were observed by electron microscope. Results The methodology for isolation and expansion of MSCs was established. The isolated MSCs were set very well in culture of DMEM-F12 (dulbecco's modified Eagle's medium-F12) supplemented with 10% selected fetal bovine serum (FBS). MSCs attached and grew as fibroblastic cells at 24 hours after initial plating and developed into visible symmetric colonies at about 36-48 hours, 7-10 days later it reached confluence while the cells were permitted to proliferate. The swine MSCs from 10 ml bone marrow aspirate showed a fibroblast-like morphology with vortex distribution in their peak propagation and were harvested (3.8±0.6)×10^6 cells after cultured three generations over 2-3 weeks. The MSCs did not differentiate spontaneously during culture expansion and maintained normal biological features. The morphology of MSCs were augmented and lengthened growing with well and closed-up arranged after chemical inducement. Muhinuclear cells and myotube structures were found more in 5,10 μmol/L 5-aza treatment groups with inducement rate were 36% and 31% respectively (no significant difference) than in 3μmol/L and control

作者信息

序号	姓名	单位	发文数	被引次数	H指数	G指数
1	程瑞	解放军总医院老年心血管病研究所	1	0	0.0	0.0