CAJ | 학술논문

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.
배경:전렬선암간세포시전렬선암복발침습적중요원인,목전연구난점재우전렬선암간세포분리기술효솔교저.목적:탐색고효지종인전렬선암PC-3급LNCap세포주분리전렬선암간세포적방법.방법:채용함혈청첩벽배양법급무혈청현부배양법배양PC-3급LNCap세포주,연후이용류식세포표면표기CD133급CD44검측량충세포재불동배양조건하가획취전렬선암간세포적비례,동시채용유도분화실험초보감정전렬선암간세포특성.결과여결론:PC-3급LNCap세포능재첨가생장인자적무혈청배양기중형성현부세포구,접충재함혈청배양기후가이유도분화위첩벽세포;무혈청배양조적CD44+/CD133+세포비례:PC-3위0.59%,LNCap위1.71%,함혈청배양조중적CD44+/CD133+세포비례:PC-3위0.32%,LNCap위0.73%,기중LNCap세포채용량충방법소획적CD44+/CD133+세포균고우PC-3소획적적세포(P < 0.05),재량충세포중무혈청현부배양화함혈청첩벽배양차이무현저성의의 (P > 0.05),단무혈청현부배양주기장,획득세포수상대교소,직접영향분선후종류간세포공능측정.인차가이증실함혈청첩벽배양LNCap세포교무혈청현부배양법경능고효쾌첩적획취전렬선암간세포.
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.