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目的体外分离培养鉴定躯干神经嵴干细胞,探讨其生物学特性,为研究周围神经再生修复提供新的细胞来源。方法取大鼠胚胎躯干部神经管,组织块法培养获取躯干神经嵴干细胞,通过形态学及免疫细胞化学观察其细胞生长、增殖、分化等特性。结果原代培养的躯干神经嵴干细胞呈梭形,其特异性标记物nestin和p75表达阳性,并有较高的增殖活性。传代培养后,在含血清培养基中,其分化细胞呈GFAP、MAP-2、α-SMA阳性。结论成功地培养了躯干神经嵴干细胞,细胞表现出多潜能干细胞特性,为研究周围神经组织工程种子细胞来源提供实验依据。
OBJECTIVE: To isolate and culture human neural crest stem cells (MSCs) in vitro and explore their biological characteristics to provide a new source of cells for the study of peripheral nerve regeneration and repair. Methods The neural tube of the trunk of the rat embryo was harvested and the neural crest stem cells of the trunk were obtained by tissue block method. The growth, proliferation and differentiation of the stem cells were observed by morphology and immunocytochemistry. Results Primary cultured neural crest stem cells were fusiform, and their specific markers nestin and p75 were positive, and had higher proliferative activity. After subculture, the differentiated cells were positive for GFAP, MAP-2 and α-SMA in serum-containing medium. Conclusion Trigeminal neural crest stem cells were successfully cultured. The cells showed the characteristics of pluripotent stem cells and provided experimental evidence for studying the source of seed cells of peripheral nerve tissue engineering.