摘要
目的 探讨人脐血CD34+ 细胞群中分离培养造血基质细胞的可行性。方法 分离人脐血CD34+ 细胞 ,采用Dexter法培养 ,对贴壁细胞进行观察和鉴定。结果 Dexter培养 9～ 1 4d(平均 1 1 .2d)开始形成基质细胞集落 ,1 5～ 2 2d(平均 1 9.6d)集落数量最多 ,培养 2 8d贴壁细胞铺满培养皿底。细胞类型以成纤维样细胞、巨噬样细胞、“小圆”类细胞为主 ;细胞化学染色 :非特异性酯酶染色 (NSE)、糖原染色 (PAS) :1 0 0 %阳性 ,过氧化物酶染色 (POX) :阴性 ,碱性磷酸酶 (ALP) :2 8%阳性 ;免疫组化染色 :CD1 0 6 :阳性 96 % ,CD2 9:93%阳性 ,CD4 4 :98%阳性 ,CD4 5 :阴性 ,CD5 0 :6 2 %阳性 ,纤维粘连蛋白 (Fn) :92 %阳性 ,层粘连蛋白(Lm) :74 %阳性 ,胶原Ⅳ :83%阳性。
Objective To study the possibility of culture of human umbilical cord blood stromal cell(HUCBSC) from cord CD34 + cells. Methods The umbilical cord blood CD34 + cells were cultured in Dexter system in order to observe the biological behavior of adherent cells in vitro. Results All cells were cultured with Dexter system. By 9 14d(at a median of 11.2d),stromal cell colonies formed and reached their maximum at 15 22d(mean 19.6d), by 28d, all adherent cells filled the basis of petri dish. Being observed from light microscope, these cells differentiated to three kinds of cells in 28d cultured:fibroblast liked cell, macrophage liked cell and small sphere cells. The ratio of these three kinds of cells was 56.8%,38%,5.5% respectively. Cytochemical : the positive rate reached 100% in NSE stain and PAS stain, ALP stain for stromal cell also manifested 35% positive, but in POX stain the result was negative. Immunohistochemistry stain: the positive rate of cord stromal cells for CD106, CD29, CD44, CD45, CD50, Fn, Lm , collagenⅣ,etc,reached 96%, 93%,98%, 68%,72%,92%,74%,83% respectively.Conclusion There are hematopoietic stromal precursor in HUCB.
出处
《重庆医学》
CAS
CSCD
2003年第10期1300-1301,1306,共3页
Chongqing medicine
基金
国家自然科学基金资助项目 (30 0 70 32 7)
关键词
CD34^+细胞
脐血
体外培养
人脐血基质细胞
CD34 + cells
human umbilical cord blood(HUCB)
culture in vitro
human umbilical cord blood stromal cell(HUCBSC)
