南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (08): 1151-.

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小鼠脂肪间充质干细胞的分离培养及其在肝内归巢

赵文秀,张磊,刘建明,尹震宇,王效民   

  • 出版日期:2013-08-20 发布日期:2013-08-20

Isolation, culture and homing to the liver of mouse adipose-derived stem cells

  • Online:2013-08-20 Published:2013-08-20

摘要: 目的建立小鼠脂肪间充质干细胞(ADSCs)分离培养的方法,并将ADSCs移植于体内,观察其致瘤性及肝脏归巢情况。
方法取皮下脂肪用0.075%Ⅰ型胶原酶消化,贴壁培养获取小鼠ADSCs,通过流式细胞技术检测其表面分子、细胞周期;并将
ADSCs注射到小鼠皮下观察其致瘤性,及尾静脉注射观察其肝脏的归巢。结果分离培养的细胞高表达CD29、CD44,不表达
CD34、CD45、CD11b、CD14;细胞周期显示G0/G1、S、G2/M 的细胞所占的比例分别为80.1%、7.9%、12%;ADSCs具有低免疫原
性,不表达CD40、CD80、CD86、MHCI 、MHCII以及PDL-1。在IFN-γ作用下,可轻度上调CD40、CD80以及PDL-1。ADSCs移
植后未见成瘤,且可在肝脏实质内多处定居。结论胶原酶消化、贴壁培养可以从小鼠脂肪中分离出高纯度的间充质干细胞,且
ADSCs移植后能在肝内定居。

Abstract: Objective To explore the method for the isolation, cultivation, and purification of adipose-derived stem cells
(ADSCs) and examine the oncogenesis and homing of ADSCs to the liver in vivo. Methods ADSCs were isolated from female
mice by digestion with 0.075% collagenase I and the morphology of the isolated cells was observed with examination of the
cell surface markers and cell cycle. BALB/c mice were injected with 1×106 ADSCs on the back to evaluate the oncogenesis of
ADSCs or with 1 × 106 ADSCs stained with 5, 6-carboxyfluorescein diacetate-succinimidyl ester (CFSE) via the tail vein to
examine the cell homing to the liver. Results The isolated ADSCs highly expressed CD29 and CD44 and were negative for
CD34, CD45, CD11b and CD14. Cell cycle distribution analysis showed cell percentages in G0/G1, S, and G2/M phases of
80.1%, 7.9%, and 12%, respectively. The ADSCs had a low immunogenicity and did not express CD40, CD80, CD86, MHCI,
MHCII or PDL-1. After stimulation with IFN-γ, the expression of CD40, CD80 and PDL-1 were up-regulated slightly in the
cells. Dorsal injection of the ADSCs did not result in any tumor formation within 1 month, and ADSCs injected via the tail vein
showed cell homing to the liver. Conclusion Murine ADSCs can be isolated and expanded effectively by collagenase digestion
and adherent culture. The isolated ADSCs can successfully reside in the liver after implantation, and thus may serve as a
promising candidate cell in stem cell therapy of liver diseases.