肠与脑来源神经干细胞的增殖和分化特性比较

The Comparative Study on the Proliferation and Differentiation of Neural Stem Cells from the Gut and Brain of Rats.

目的探索肠神经干细胞(ENSCs)的增殖和分化特性,寻找神经干细胞移植治疗肠道神经元缺陷性疾病的合适干细胞源。方法采用神经干细胞培养和分离技术,对来自同一大鼠肠道及大脑的神经干细胞(CNS-NSCs)进行体外培养,通过神经球生长直接观察法、CCK-8细胞增殖测定法、流式细胞仪测定,比较研究两者的生长和增殖情况及成熟分化特性。结果与CNS-NSCs相比,ENSCs神经球增殖速度较慢;ENSCs神经球表面常有长突起,并相互连接成网络状,显示早期成熟分化的迹象;CCK-8法测得ENSCs的吸光度明显低于CNS-NSCs(P<0.05),提示其增殖速度慢于CNS-NSCs。流式细胞仪测得EN-SCs和CNS-NSCs的TUJ-1阳性细胞比例分别为12.3%和11.4%(P>0.05)、GFAP阳性细胞比例分别为21.7%和22.5%(P>0.05)、Nestin阳性细胞比例分别为10.8%和10.8%(P>0.05)。结论 ENSCs与CNS-NSCs具有类似的增殖、分化特性,但ENSCs增殖速度较慢,易于成熟分化。

Objective To explore the characteristics of proliferation and differentiation of enteric neural stem cells （ENSCs） , and look for the suitable source of neural stem cells for transplantation treatment of developmental defects of enteric neuron. Methods We used the culture medium and isolation technique adopted in our laboratory to culture in vitro enteric neural stem cells （ENSCs） from gut and central nervous system - derived neural stem cells （ CNS - NSCs） from brain obtained from the same embryonic rat. The state of growth, the characteristics of proliferation and differentiation of both neural stem cells were compared by direct observation of neuro - sphere growth, CCK - 8 method and flow cytometry. Results Compared to the CNS - NSCs, the proliferation of ENSCs neurosphere was much slower. Some long dendrites often grew from the surface of the ENSCs neurosphere, and connected each other and formed a network, which seemed to differentiation in early stage. The growth rate of the ENSCs was much lower than that of CNS - NSCs（P 〈 0.05 ） by CCK -8 method. The percentage of TUJ- 1 positive cells of ENSCs and CNS- NSCs were 12.3% and 11.4% respectively （P 〉 0.05 ）by flow cytometry. The percentage of GFAP positive cells of ENSCs and CNS - NSCs were 21.7% and 22.5% respectively （ P 〉 0.05 ）. The percentage of Nestin positive cells of ENSCs and CNS - NSCs were 10.8% and 10.8% respectively（P 〉 0.05 ）. Conclusion ENSCs and CNS -NSCs have similar biological characteristics of proliferation and differentiation,but ENSCs are much slower in proliferation and easier to differentiate and maturate.