Liang Peng, Zhao Shiguang, Kawamoto Keiji, Jin Lianhong, Liu Enzhong
Department of Neurosurgery, First Medical College of Harbin Medical University, Harbin 150001, PR China.
Hum Cell. 2003 Sep;16(3):151-6. doi: 10.1111/j.1749-0774.2003.tb00147.x.
To set up long-term in vitro culture system of the human neural stem cells (hNSC) and to study their biological properties.
Human fetuses aged about 20 weeks following spontaneous abortion were adopted. A serum-free medium containing basic fibroblast growth factor and epidermal growth factor was used to make the hNSCs divide continuously in the culture. The growth curve of continually passaged cells was examined. The effects of long-term culture on the cell cycle, cell differentiation were analyzed. The cell cycles of these cells were analyzed using flow cytometry.
The cells from the human embryonic cortical tissue could be maintained and propagated in the presence of growth factors. Neurospheres were generated continually. Only one month after the primary culture, the precursors could be effectively discarded. The cells could be cultured for ten months. The cells had an exponential, consistent growth. The cell cycle analysis indicated that the hNSCs maintained remarkable proliferation. Upon differentiation, the hNSCs gave rise to mature cells. The multi-lineage potential of differentiation after different passages remained unchanged.
The hNSCs isolated from the human embryonic tissues retained their biological features after long-term culture in vitro.
建立人神经干细胞（hNSC）的长期体外培养体系并研究其生物学特性。
采用自然流产的约20周龄人类胎儿。使用含有碱性成纤维细胞生长因子和表皮生长因子的无血清培养基使hNSC在培养中持续分裂。检测连续传代细胞的生长曲线。分析长期培养对细胞周期、细胞分化的影响。使用流式细胞术分析这些细胞的细胞周期。
人胚胎皮质组织来源的细胞在生长因子存在的情况下能够维持并增殖。持续产生神经球。原代培养仅1个月后，前体细胞就能被有效去除。细胞可培养10个月。细胞呈指数性、一致性生长。细胞周期分析表明hNSC保持着显著的增殖能力。在分化时，hNSC分化为成熟细胞。不同传代后多谱系分化潜能保持不变。
从人胚胎组织分离的hNSC在体外长期培养后仍保留其生物学特性。
