中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (6): 1011-1014.doi: 10.3969/j.issn.1673-8225.2012.06.014

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

前列腺癌干细胞分离方法的对比及筛选********★

李奎庆，许可慰，周邦奋，范新兰，董  文，张彩霞，毕良宽   

  1. 中山大学孙逸仙纪念医院泌尿外科，广东省广州市  510120
  • 收稿日期:2011-07-14 修回日期:2011-08-10 出版日期:2012-02-05 发布日期:2012-02-05
  • 通讯作者: 许可慰，硕士生导师，副教授，中山大学孙逸仙纪念医院泌尿外科，广东省广州市 510120
  • 作者简介:李奎庆★，男，1985年生，中山大学孙逸仙纪念医院在读硕士，主要从事前列腺癌干细胞研究。
  • 基金资助:

    国家自然科学基金项目(81001138)， 国家自然科学基金青年基金项目(30901488)，教育部新教师基金(200805581124)，广东省自然科学基金(06021283、10151008901000024)，广东省科技计划项目(2008B0303 01078)，广东省自然科学基金博士启动基金项目(9451008901003001)，广东省医学科学技术研究基金项目(A2008189)。

Comparison and screening of prostate cancer stem cells isolation methods********★

Li Kui-qing, Xu Ke-wei, Zhou Bang-fen, Fan Xin-lan, Dong Wen, Zhang Cai-xia, Bi Liang-kuan   

  1. Department of Urinary Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, Guangdong Province, China
  • Received:2011-07-14 Revised:2011-08-10 Online:2012-02-05 Published:2012-02-05
  • Contact: Xu Ke-wei, Master, Associate professor, Department of Urinary Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, Guangdong Province, China xkw2005@163.com
  • About author:Li Kui-qing★, Studying for master’s degree, Department of Urinary Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, Guangdong Province, China kuiqing04@yahoo.com.cn
  • Supported by:

    the National Natural Science Foundation of China, No. 81001138*; the National Natural Science Youth Foundation of China, No. 30901488*; the New Teacher Foundation of Ministry of Education, No. 200805581124*; the Natural Science Foundation of Guangdong Province, No. 06021283*, 1015100 8901000024*; the Science and Technology Development Program of Guangdong Province, No. 2008B030301078*; the Natural Science Foundation for Doctors of Guangdong Province, No. 94510 08901003001*; the Medical Science and Technology Research Foundation of Guangdong Province, No. A2008189*

PDF

375

被引次数

5

摘要：

背景：前列腺癌干细胞是前列腺癌复发侵袭的重要原因，目前研究难点在于前列腺癌干细胞分离技术效率较低。
目的：探索高效地从人前列腺癌PC-3及LNCap细胞株分离前列腺癌干细胞的方法。
方法：采用含血清贴壁培养法及无血清悬浮培养法培养PC-3及LNCap细胞株，然后利用流式细胞表面标记CD133及CD44检测两种细胞在不同培养条件下可获取前列腺癌干细胞的比例，同时采用诱导分化实验初步鉴定前列腺癌干细胞特性。
结果与结论：PC-3及LNCap细胞能在添加生长因子的无血清培养基中形成悬浮细胞球，接种在含血清培养基后可以诱导分化为贴壁细胞；无血清培养组的CD44+/CD133+细胞比例：PC-3为0.59%，LNCap为1.71%，含血清培养组中的CD44+/CD133+细胞比例：PC-3为0.32%，LNCap为0.73%，其中LNCap细胞采用两种方法所获的CD44+/CD133+细胞均高于PC-3所获的的细胞(P < 0.05)，在两种细胞中无血清悬浮培养和含血清贴壁培养差异无显著性意义 (P > 0.05)，但无血清悬浮培养周期长，获得细胞数相对较少，直接影响分选后肿瘤干细胞功能测定。因此可以证实含血清贴壁培养LNCap细胞较无血清悬浮培养法更能高效快捷的获取前列腺癌干细胞。

关键词: 前列腺癌干细胞, 无血清培养基, 血清培养基, 悬浮培养, 流式分选

Abstract:

BACKGROUND: Prostate cancer stem cell is an important reason for the invasion and recurrence of prostatic carcinoma. However, separation efficiency of prostate cancer stem cells is very low.
OBJECTIVE: To explore the efficient method for isolating and identifying the prostate cancer stem cells from human prostatic carcinoma cell lines PC-3 and LNCap.
METHODS: Prostate cancer cell lines PC-3 and LNCap were cultured in serum free medium (SFM) and serum supplemented medium (SSM), respectively. The percentage of prostate cancer stem cells from different medium was detected by flow cytometry through markers CD133 and CD44, and the properties of prostate cancer stem cells were preliminarily identified using inducing differentiation experiments.
RESULTS AND CONCLUSION: PC-3 and LNCap formed sphere cells in SFM, which can be induced into adherent cells after culture in SSM. Higher percentage of CD44+/CD133+cells was obtained from LNCap cells (1.71%; 0.73%) than PC-3 cells (0.59%; 0.32%) in both SFM and SSM. The number of CD44+/CD133+ LNCap cells was more than PC-3 using both methods (P < 0.05), but the efficiency of SFM and SSM had no statistical significance (P > 0.05). However, the culture cycle was longer and number of obtained cells was less by SFM culture, directly influencing functional determination of prostate cancer stem cells. Compared with suspension culture method with SFM, SSM is more convenient and effective in isolating prostate cancer stem cells from LNCap cells.

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