Peiris Tanuja Harshani, García-Ojeda Marcos E, Oviedo Néstor J
Department of Molecular and Cell Biology, School of Natural Sciences University of California Merced California 95343 USA; Quantitative and Systems Biology Graduate Program University of California Merced California 95343 USA.
Department of Molecular and Cell Biology, School of Natural Sciences University of California Merced California 95343 USA; Quantitative and Systems Biology Graduate Program University of California Merced California 95343 USA; Health Sciences Research Institute University of California Merced California 95343 USA.
Regeneration (Oxf). 2016 Mar 16;3(2):123-35. doi: 10.1002/reg2.53. eCollection 2016 Apr.
Planarians possess remarkable stem cell populations that continuously support cellular turnover and are instrumental in the regeneration of tissues upon injury. Cellular turnover and tissue regeneration in planarians rely on the proper integration of local and systemic signals that regulate cell proliferation and cell death. Thus, understanding the signals controlling cellular proliferation and cell death in planarians could provide valuable insights for maintenance of adult body homeostasis and the biology of regeneration. Flow cytometry techniques have been utilized widely to identify, isolate, and characterize planarian stem cell populations. We developed alternative flow cytometry strategies that reduce the number of reagents and the time of sample preparation to analyze stem cells and cell death in planarians. The sensitivity of these methods is validated with functional studies using RNA interference and treatment with γ irradiation or stressful conditions that are known to trigger cell death. Altogether, we provide a community resource intended to minimize adverse effects during ex vivo studies of stem cells and cell death in planarians.
涡虫拥有显著的干细胞群体，这些干细胞群体持续支持细胞更新，并在损伤后组织的再生中发挥重要作用。涡虫的细胞更新和组织再生依赖于调节细胞增殖和细胞死亡的局部和全身信号的适当整合。因此，了解控制涡虫细胞增殖和细胞死亡的信号，可为维持成体体内稳态和再生生物学提供有价值的见解。流式细胞术技术已被广泛用于鉴定、分离和表征涡虫干细胞群体。我们开发了替代的流式细胞术策略，减少了试剂数量和样品制备时间，以分析涡虫中的干细胞和细胞死亡。这些方法的灵敏度通过使用RNA干扰以及用已知会触发细胞死亡的γ射线照射或应激条件进行的功能研究得到验证。总之，我们提供了一种旨在尽量减少涡虫干细胞和细胞死亡体外研究期间不利影响的社区资源。
