自制免疫磁珠在前列腺肿瘤组织干细胞提取中的应用

Extraction of prostate cancer stem cells using self-designed magnetic beads

背景:有效分离干细胞是前列腺癌发生发展研究的基础,目前的主流方法是免疫磁珠分选。目的:应用自制的免疫磁珠分选前列腺肿瘤组织中的CD133^+/CD44^+细胞,并进一步培养和传代进行免疫学鉴定,以确定免疫磁珠分选效果。方法:应用独创的磁珠制备免疫磁珠,分选肿瘤组织中细胞膜表达CD133和CD44的细胞。观察分选细胞无血清培养成球情况,及传代后其在形态学、增殖能力方面与标准肿瘤细胞株的差异,并用免疫荧光法检测其特异性抗体的表达。结果与结论:制备的免疫磁珠粒径小、分选效果佳。分选的细胞成球效果好,培养传代后的细胞依旧明显表达CD133和CD44抗原。分选的细胞非诱导培养后可见多种形态,生长旺盛。转化生长因子β诱导培养后细胞形态较单一且生长缓慢。两组细胞与标准肿瘤细胞株培养后的增殖情况差异均有显著性意义(P<0.05)。结果表明应用自制免疫磁珠从肿瘤细胞中分选的CD133^+/CD44^+细胞经过初步形态及功能鉴定具有干细胞的特性,可为前列腺癌干细胞的提取及进一步培养研究奠定基础。

BACKGROUND：Effective sorting of prostate cancer stem cells is the basis of experimental studies in prostate cancer developing.The most common sorting method is magnetic-activated cell sorting.OBJECTIVE：To separate CD133^＋/CD44^＋ cells in prostate cancer tissues using self-designed magnetic beads followed by culture,passage and immunological identification.METHODS：Self-designed magnetic microspheres were applied to establish immunomagnetic beads to sort CD133^＋/CD44^＋ cells in prostate cancer tissues.The sorted cells were cultured in serum-free medium.The sphere formation,cell morphology,and proliferation ability after cell passage were statistically compared between the sorted cells and the normal tumor cell lines.Immunofluorescence detection was performed to detect the expression of specific antibodies.RESULTS AND CONCLUSION：Self-designed immunomagnetic beads had small diameter and a high-sorting effect.The sorted cells possessed a high capacity of microsphere formation.After cell culture and passage,the cells highly expressed CD133 and CD44 antigens.The sorted cells with no induction had varying shapes and grew vigorously.After induction with transforming growth factor-β,the cultured cells were noted to have a single shape and grow slowly.The cell proliferation ability of sorted cells in these two groups differed significantly from that of the normal cancer cell lines（both P〈0.05）.In conclusion,the CD133^＋/CD44^＋ cells sorted from prostate tumor cells possessed cell morphology and function characteristics of stem cells which can provide a basis for extraction and culture of prostate cancer stem cells.