Institute of Hematology, Department of Hematology, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University; The Lab of Aging Research, State Key Laboratory of Biotherapy, National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University.
Institute of Hematology, Department of Hematology, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University.
J Vis Exp. 2024 Aug 23(210). doi: 10.3791/67012.
The side population (SP) cells are identified through Hoechst 33342 staining and analyzed using flow cytometry (FCM). The Hoechst SP method is utilized for the isolation of stem cells based on the dye efflux properties of ATP-binding cassette (ABC) transporters. The method was initially employed for the identification and isolation of hematopoietic stem cells (HSCs), but it has now evolved to primarily focus on the identification and isolation of cancer stem cells (CSCs). The traditional detection method of FCM uses a 355 nm laser to excite the dye to detect SP cells. Through this study, we have successfully identified alternative approaches for dye excitation that can effectively replace the detection of SP cells using a 355 nm laser. This is achieved through the utilization of high-power 375 nm or 405 nm lasers. This allows us to exercise enhanced selectivity in the detection of SP cells rather than being solely limited to the 355 nm laser flow cytometry.
侧群 (SP) 细胞通过 Hoechst 33342 染色进行鉴定，并通过流式细胞术 (FCM) 进行分析。Hoechst SP 方法基于三磷酸腺苷结合盒 (ABC) 转运蛋白的染料外排特性，用于分离干细胞。该方法最初用于鉴定和分离造血干细胞 (HSCs)，但现在已发展为主要用于鉴定和分离癌症干细胞 (CSCs)。FCM 的传统检测方法使用 355nm 激光激发染料以检测 SP 细胞。通过这项研究，我们成功地找到了替代染料激发的方法，可以有效地替代使用 355nm 激光检测 SP 细胞。这是通过使用高功率 375nm 或 405nm 激光来实现的。这使我们能够在检测 SP 细胞时具有更高的选择性，而不仅仅局限于 355nm 激光流式细胞术。
