前列腺癌干细胞分离方法的对比及筛选

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。

Comparison and screening of prostate cancer stem cells isolation methods

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.