CAJ | 학술논문

目的研究体外培养条件下,脑微血管内皮细胞(brain microvascular endothelial cells,BMECs)对脑肿瘤干细胞(braintumor stem cells,BTSCs)增殖、自我更新及分化功能的影响.方法应用Transwell小室建立BMECs与BTSCs共培养模型,并建立对照组,共培养14 d后,测量肿瘤球(brain tumor sphere,BTS)直径大小,检测BTSCs增殖曲线,测定单细胞克隆形成率.在含血清的培养基中培养10 d后,行CD133、Nestin、GFAP、DAPI免疫荧光染色,计数两组的CD133、Nestin、GFAP染色的细胞数及同一视野的DAPI染色细胞核数,观察两组之间的差异.结果共培养组BTS直径是对照组的5.05倍,增殖快,共培养组有61.4%能形成下一代BTSCs,而对照组为39.0%.在含血清的培养基中培养10 d后,共培养组的GFAP阳性率较对照组低,而CD133、Nestin阳性率较对照组高.结论 BMECs能够促进BTSCs增殖、自我更新能力,保持BTSCs未分化状态.
목적 연구체외배양조건하,뇌미혈관내피세포(brain microvascular endothelial cells,BMECs)대뇌종류간세포(braintumor stem cells,BTSCs)증식、자아경신급분화공능적영향.방법 응용Transwell소실건립BMECs여BTSCs공배양모형,병건립대조조,공배양14 d후,측량종류구(brain tumor sphere,BTS)직경대소,검측BTSCs증식곡선,측정단세포극륭형성솔.재함혈청적배양기중배양10 d후,행CD133、Nestin、GFAP、DAPI면역형광염색,계수량조적CD133、Nestin、GFAP염색적세포수급동일시야적DAPI염색세포핵수,관찰량조지간적차이.결과 공배양조BTS직경시대조조적5.05배,증식쾌,공배양조유61.4%능형성하일대BTSCs,이대조조위39.0%.재함혈청적배양기중배양10 d후,공배양조적GFAP양성솔교대조조저,이CD133、Nestin양성솔교대조조고.결론 BMECs능구촉진BTSCs증식、자아경신능력,보지BTSCs미분화상태.
Objective To investigate the effect of the brain microvascular endothelial cells (BMECs) on proliferation, self - renewal and differentiation of the brain tumor stem cells (BTSCs) in vitro. Method To establish a co - cultural system between BMECs and BTSCs on Transwell inserts in vitro. While the control group was preformed. The co - culture was maintained for 14 days. Then the size of brain tumor spheres (BTS) and the growth curves were determined. The single - colone formed rates of two groups were determined. The BTSCs of two group were cultured in DMEM/F12 containing 10% fetal calf groups were determined.The BTSCs of two group were cultured in DMEM/F12 containing 10% fetal calf serum for 10 days. Then CD133,Nestin,GFAP and DAPI immunofluorescence staining were made. CD133, Nestin,GFAP positive rate of two groups were calculated separately. The differences between the two groups were observed. Results BTS that were co - cultured with BMECs increased in size up to 5. 05 times larger than those grown in control group. There were 61. 4% BTSCs of the co - cultural group generate next tumor spheres. But that of the control group was 39. 0%. After BTSCs were cultured in the culture medium with 10% fetal bovine serum for 10 days,we found that GFAP - positive rate of the co - cultural group was lower than that of the control group. While CD 133 -positive and Nestin - positive rate of the co-cultural group was higher than that of the control group. Conclusions Brain microvascular endothelial cells could promote the proliferation and self - renewal and maintain the undifferentiated state of brain tumor stem cells.