卵黄囊干细胞向粒-单系造血祖细胞的定向诱导分化

目的：观察卵黄囊干细胞向粒-单系造血祖细胞（CFU-GM）的定向诱导分化，稳定和优化检测卵黄囊CFU-GM的方法。方法：应用体外琼脂培养法，观察多种因素对小鼠卵黄囊干细胞形成粒-单系造血祖细胞集落形成单位的影响；应用染色压片法对所生成的集落性质作鉴定。结果：植入不同数量的E7.5~8.5 d的卵黄囊干细胞与CFU-GM生成数量之间呈高度正相关；在接种相同数量的卵黄囊细胞数的情况下，DMEM培养体系生成的CFU-GM数明显多于IMDM培养体系生成的集落数（P<0.01）；马血清（HS）组优于胎牛血清（FBS）组（P<0.01）；与GM-CSF相比，WEHI-3条件培养液（W3-CM）能明显促进卵黄囊CFU-GM的生成（P<0.01）。采用完整琼脂凝块压片法对卵黄囊细胞所形成的集落进行鉴定表明为粒-单系细胞集落。结论：卵黄囊干细胞具有向CFU-GM分化的能力；卵黄囊干细胞CFU-GM诱导体系中各组分变化均可影响CFU-GM的集落生成率；以DMEM，GM-CSF或W3-CM，HS为组分的体系可稳定地检测卵黄囊细胞CFU-GM的集落生成率。

Induction of committed differentiation from yolk sac stem cells to CFU-GM

OBJECTIVE: To set up a stable technology for the growth of yolk sac CFU-GM. METHODS: The effects of various inductive factors on the growth of yolk sac CFU-GM and the characteristic of colonies were investigated. In vitro agar culture method was used in this study. RESULTS: We found close relationship between the number of E7.5 - E8.5 yolk sac cells planted and the number of the formation of CFU-GM colonies. The number of the formation of CFU-GM colonies in DMEM culture system was higher than that in the IMDM culture system with the same number of yolk sac cells planted. The effect of HS culture system on the formation of CFU-GM colonies was better than that of FBS culture system. In contrast with GM-CSF, WEHI-3-CM had much more significant CFU-GM growth promoting effect (4.14 times improved, P < 0.01). The colonies derived from yolk sac cells in this study were CFU-GM colonies by in situ fixation and H. E staining. CONCLUSION: The committed differentiation from yolk sac stem cells to CFU-GM is affected by many factors. The combination of DMEM, HS, GM-CSF or WEHI-3-CM in culture system is optimal for yolk sac CFU-GM growth.