中国组织工程研究

中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (1): 36-41.doi: 10.3969/j.issn.2095-4344.2016.01.007

• 肿瘤干细胞 cancer stem cells • 上一篇    下一篇

自制免疫磁珠在前列腺肿瘤组织干细胞提取中的应用

龚 锐1，李胜英2，霍志霞3，丁 浩4，孙二琳4   

  1. 1天津医科大学第二医院药学部，天津市 300211；2天津市儿童医院检验科，天津市 300070；3天津大学药物科学与技术学院，天津市 300050；4天津医科大学第二医院泌尿外科，天津市泌尿外科研究所，天津市 300211
  • 收稿日期:2015-10-26 出版日期:2016-01-01 发布日期:2016-01-01
  • 作者简介:龚锐，女，1975年生，山东省掖县人，硕士，主管药师，主要从事药物分析方面的研究。
  • 基金资助:
    天津市卫生和计划生育委员会科技基金(2014KZ093)

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

Gong Rui1, Li Sheng-ying2, Huo Zhi-xia3, Ding Hao4, Sun Er-lin4   

  1. 1Pharmaceutical Department, the Second Hospital of Tianjin Medical University, Tianjin 300211, China; 2Clinical Laboratory, Tianjin Children’s Hospital, Tianjin 300070, China; 3School of Pharmacology Science and Technology, Tianjin University, Tianjin 300050, China; 4Department of Urology, the Second Hospital of Tianjin Medical University, Tianjin Institute of Urology, Tianjin 300211, China
  • Received:2015-10-26 Online:2016-01-01 Published:2016-01-01
  • About author:Gong Rui, Master, Pharmacist in charge, Pharmaceutical Department, the Second Hospital of Tianjin Medical University, Tianjin 300211, China
  • Supported by:

    the Science and Technology Fund of Tianjin Public Health Bureau, No. 2014KZ093

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摘要：

文章快速阅读：

 

文题释义：
免疫磁珠细胞分选技术：免疫磁珠细胞分选方法可以在几分钟内从复杂的细胞混合物中分离出很高纯度的目的细胞。免疫磁珠的体积很小，不会对细胞造成机械性压力，且孵育时间短、操作过程快，阳性分选出的细胞可立即用于分析和随后的实验。
肿瘤干细胞的分选：肿瘤干细胞在肿瘤组织中只占0.02%-0.1%，但其具有自我更新能力并能产生异质性肿瘤细胞，分选高纯度的肿瘤干细胞在肿瘤组织的生长、转移和复发研究中具有重要意义。

 

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。

关键词: 干细胞, 肿瘤干细胞, 免疫磁珠细胞分选, CD133, CD44

Abstract:

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.

 

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