人脐血单个核细胞体外分化为神经细胞  

The preliminary study on in vitro differentiation of human umbilical cord blood cells into neural cells

作　　者：赵宗茂[1] 卢士红[2] 张庆俊[1] 刘海英[1] 杨仁池[2] 李洪钧[2] 蔡英林[2] 韩忠朝[2] 

机构地区：[1]河北医科大学第二医院神经外科 [2]中国医学科学院,中国协和医科大学血液学研究所,实验血液学国家重点实验室

出　　处：《中华血液学杂志》2003年第9期484-487,共4页Chinese Journal of Hematology

基　　金：国家 973项目 (0 0 1CB5 10 1);攀登计划 (95 专 10 )资助项目

摘　　要：目的　探讨核转录因子 (NF) κB配体受体激动因子 (receptoractivatorofNF KappaBligand ,RANKL)和脑源性神经营养因子 (brain derivedneurotrophicfactor,BDNF)体外诱导人脐血细胞 (humanum bilicalcordbloodcells,HUCBC)分化为神经元和神经胶质细胞的可行性。方法　采集足月妊娠、正常分娩的新鲜脐血 ,取单个核细胞 (MNC) ,利用人可溶性NF κB配体受体激动因子 (sRANKL)、BDNF作为分化诱导因子进行体外诱导 ,维甲酸 (RA)诱导分化作为对照 ,倒置显微镜动态观察细胞生长、分化情况 ;培养 10d后 ,应用免疫细胞化学染色法检测培养细胞的胶质纤维酸性蛋白 (GFAP)和神经元特异核蛋白 (NeuN)表达情况 ,进行细胞性质鉴定。结果　诱导分化 10d后 ,RANKL、BDNF和BDNF +RANKL组的NeuN阳性细胞数分别为 (97.0± 13.5 )个 ,(85 .0± 5 .6 )个 ,(16 7.0± 19 7)个 ,分别是对照组 [(5 5 .7±8.5 )个 ]的 1.7,1.5 ,3.0倍 ,GFAP阳性细胞数分别为 114 .7± 18.0 ,2 33.3± 2 1.7,2 89.0± 2 4 .9,分别是对照组的 1.4 ,2 .9,3.6倍。RANKL向神经元方向诱导分化的作用与BDNF相当 ,向神经胶质细胞分化的作用不及BDNF。RANKL和BDNF对HUCBC向神经细胞分化具有协同作用 ,二者联合应用明显促进HUCBC向神经元和神经胶质细胞分化?Objective To explore the feasibility of in vitro differentiation of human umbilical cord blood cells (HUCBC) into neural cells induced by receptor activator of NF-KappaB ligand (RANKL) and brain-derived neurotrophic factor(BDNF).Methods Normal fresh HUCBC were cultured as the following:①Control group cultured by differentiation medium only; ②BDNF group, cultured by differentiation medium + BDNF ;③RANKL group, cultured by differentiation medium + human soluble RANKL(sRANKL); ④BDNF+ RANKL group, cultured by differentiation medium + BDNF and sRANKL.Cultured cells were observed with invert microscope. After ten-days culture, the expression of glial fibrillary acidic protein (GFAP) and neuron-specific nuclear protein (NeuN) of the cultured cells were detected by immunocytochemical staining.Results After 10 day's culture,the NeuN positive cells were (97.0±13.5),(85.0±5.6),(167.0±19.7) in RANKL, BDNF and BDNF+RANKL groups, respectively, with 1.7, 1.5, 3.0 fold in crease than that of control (55.7±8.5),the GFAP positive cells were (114.7±18.0),(233.3±21.7),(289.0±24.7),respectively,with 1.4,2.9,3.6 fold increase compared with the control group.The differentiation ratio of neurons in RANKL group was similar to that of the BDNF group, but the differentiation ratio of glial cells was lower than that in the BDNF group. In the RANKL + BDNF group, the differentiation of HUCBC into neurons and glial cells were enhanced obviously, the differentiated neural cells were typical with longer axons and dendrites. Conclusion RANKL and BDNF could induce HUCBC into neurons and glial cells, and they have synergistic effect on the induced differentiation. It is hopeful that HUCBC might be an source of stem cells for the treatment of central nervous system injury.

关 键 词：人脐血单个核细胞 细胞分化 神经细胞 核转录因子-κB配体受体激动因子 脑源性神经营养因子 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]