不同年龄段人骨髓间充质干细胞生物学特性的研究  

Age-related Biological Characteristics of Human Bone Marrow Mesenchymal Stem Cells from Different Age Donors

作　　者：黄科[1] 周敦华[1] 黄绍良[1] 梁舜华[1] 

机构地区：[1]中山大学附属二院儿科造血干细胞移植中心,广州510120

出　　处：《中国实验血液学杂志》2005年第6期1049-1053,共5页Journal of Experimental Hematology

基　　金：卫生部临床学科重点项目[2004]468

摘　　要：本研究通过对不同年龄段人骨髓间充质干细胞(mesenchymalstemcell,MSC)的研究探讨MSC生物学特点与年龄的关系,从而为临床寻找合适供体,迅速获取所需MSC提供实验依据。骨髓供体按年龄分为4组:A组(胚胎)、B组(0-20岁)、C组(20-40岁)和D组(40岁以上)。观察各组骨髓MSC的生长、增殖、分化特点;用流式细胞仪检测细胞表面标志,用ELISA方法检测培养上清造血相关因子的水平;进行核型分析及成瘤试验;评价不同年龄段供体骨髓MSC在临床造血干细胞移植中的应用价值。结果显示:各组骨髓均可培养出MSC,各组MSC表面标志无显著差异;各组MSC均具有向脂肪细胞和成骨细胞分化的能力;原代培养B组骨髓MSC含量较多,贴壁时间早,传代时间短,P0至P1时间为5.5天,传至P10需33天,8×106MNC培养至P10时MSC数达(5.19±2.15)×1010;A、C、D组P0至P1时间分别为15、7和13天,传至P10分别需50、60和72天,8×106MNC培养传代至P10时MSC数分别为(4.98±2.08)×1010、(1.86±0.47)×1010、(0.64±0.22)×1010。A组MSC较细长,细胞融合后生长无接触抑制,P15增殖速度开始减缓;B、C、D组MSC形态相似,有生长接触抑制B组P10开始增殖减缓,C、D组P8开始增殖减缓;B组MSC培养上清中SCF、FLT3L、IL6及SDF1水平高于其它各组。各组间细胞的增殖指数无显著差异。核型分析均为正常核型,成瘤实验为阴性。结论:MSC增殖生长特性与年龄密切相关;根据临床造血干细胞移植的需要,0-20岁年龄组骨髓是短时间内获取足够细胞数的理想供体,分泌HGFs水平亦占优势;0-20岁组骨髓MSC的生物学特性优于其他各组,可以作为MSC的供源。The aim of this study was to investigate the biological characteristics of human bone marrow mesenchymal stem cells from bone marrow of different age donors. The experiments were divided into four groups by donors age, group A represented MSC derived from fetal bone marrow, group B represented MSC derived from bone marrow of 0 - 20 years old donors, group C represented MSC derived from bone marrow of 20 - 40 years old donors and group D represented MSC derived from bone marrow of donors older than 40. The growth, purification, proliferation and multipotential abilities of MSC in 4 groups were observed and their immunophenotypes were determined by flow-cytometry. The level of cytokines （ IL-6, SCF, FLT-3L, SDF-1 and TGF-β1 ） were assayed by ELISA method. Cell cycles were analyzed to show the proliferation index （PrI）. MSCs derived from bone marrow of 4 groups were injected subcutaneous into NOD/SCID mouse to observe the safety. The results showed that different age donors bone marrow all gave rise to MSC. These cells were similar in morphology, antigenic phenotype, differentiation potential and cell cycle. The primary culture time of group B was shorter than other groups. The duration of passage 1 （ P1 ） was 5.5 days, and the duration of P10 was 33 days, after P10 culture, （5.19±2.15） × 10^10 MSCs were obtained from 8 × 10^6MNC of this group. The primary culture time of groups A, C, D were longer, the duration of P1 were 15, 7 and 13 days for group A, C and D respectively, and the duration of P10 was 50, 60 and 72 days for group A, C and D, respectively. After P10 culture, （4.98 ± 2.08） × 10^10, （ 1.86 ± 0.47 ） × 10^10, （0.64 ± 0.22） × 10^10 MSCs were obtained from 8 ± 10^6 MNC of group A, C and D respectively. The morphology of MSC of group A was longer and slender. The ability of expansion decreased after P15 for A group, P10 for B group and P8 for C and D groups. The levels of SCF, FLT3-L, IL-6 and SDF- 1 in group B were higher than other groups. Karyotype analysis showed

关 键 词：间充质干细胞 骨髓 造血干细胞 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]