摘要
目的探讨MCF-7细胞、BT474细胞和MDA-MB-231细胞微球体形成的影响因素,并检测其CD44+CD24-/low的表达比例,明确富集干细胞的方法,以获得稳定数量的干细胞。方法将MCF-7、BT474和MDA-MB-231乳腺癌细胞进行微球体培养,取培养第7天的微球体细胞分别进行干细胞表型的FCM检测分析,判断干细胞富集的程度。结果 MCF-7、BT474和MDA-MB-231细胞形成微球体的效率分别为(2.40±0.20)%、(1.60±0.10)%和(0.60±0.05)%。在通常的干细胞培养液中MDA-MB-231细胞很难聚团生长。但是如果去除胰岛素,MDA-MB-231细胞则聚团生长。单层培养细胞中MCF-7、BT474和MDA-MB-231的CD44+CD24-/low的比例为(1.00±0.20)%、(0.30±0.05)%和(37.00±1.80)%,微球体细胞中CD44+CD24-/low的比例分别为(8.10±0.40)%、(0.60±0.20)%和(93.80±2.10)%,乳腺癌细胞中表达CD44+CD24-/low细胞表型的比例明显低于同种细胞培养的微球体(P<0.01)。结论通过微球体培养,富集了肿瘤干细胞,但是不同乳腺癌细胞来源的微球体中干细胞所占的比例差异很大,MDA-MB-231细胞最高。
Objective In order to acquire stable amount of breast cancer stem cells(BCSCs),we try to find the exact way of enrichment of BCSCs by investigating the influence factors on the formation of mammospheres from MCF-7,BT474 and MDA-MB-231 cells,and detecting the propotion of CD44+CD24-/low among them.Methods MCF-7,BT474 and MDA-MB-231 cells were cultured in suspension to generate mammospheres.After 7 days of cultivation,the proportion of CD44+CD24-/lowcells was analyzed in monolayer culture cells and mammosphere cells by flow cytometry(FCM) in order to see the extent of enrichment of BCSCs.Results The mammosphere formation efficiency(MFE) of MCF-7,BT474 and MDA-MB-231 cells were(2.40±0.20)%,(1.60±0.10)% and(0.60±0.05)%.The formation of mammospheres from MDA-MB-231 cells was observed without insulin but hardly observed with insulin in current stem cell suspension.FCM analysis indicated that the propotion of CD44+CD24-/low expression in MCF-7,BT474 and MDA-MB-231 monolayer culture cells were(1.00±0.20)%,(0.30±0.05)% and(37.00±1.80)%,compared with(8.10±0.40)%,(0.60±0.20)% and(93.80±2.10)% of the propotion of CD44+CD24-/low expression in the mammosphere cells.The propotion of CD44+CD24-/low expression in the mammosphere cells was much higher than that in the breast cancer cells(P<0.01).Conclusions We could get enrichment of BCSCs via cultivation of mammospeheres,while the propotion of CD44+CD24-/low expression in mammosphere cells generated from different breast cancer cells are diverse greatly.Compared with others,mammosphere cells from MDA-MB-231 contained highest propotion of BCSCs.
出处
《中华临床医师杂志（电子版）》
CAS
2011年第10期2856-2863,共8页
Chinese Journal of Clinicians(Electronic Edition)
关键词
乳腺肿瘤
肿瘤干细胞
微球体
Breast neoplasms
Neoplastic stem cells
Microspheres
