Rosental Benyamin, Kozhekbaeva Zhanna, Fernhoff Nathaniel, Tsai Jonathan M, Traylor-Knowles Nikki
Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, 94305, USA.
Department of Pathology, Hopkins Marine Station, Stanford University, 120 Ocean View Blvd, Pacific Grove, CA, 93950, USA.
BMC Cell Biol. 2017 Aug 29;18(1):30. doi: 10.1186/s12860-017-0146-8.
Generalized methods for understanding the cell biology of non-model species are quite rare, yet very much needed. In order to address this issue, we have modified a technique traditionally used in the biomedical field for ecological and evolutionary research. Fluorescent activated cell sorting (FACS) is often used for sorting and identifying cell populations. In this study, we developed a method to identify and isolate different cell populations in corals and other cnidarians.
Using fluorescence-activated cell sorting (FACS), coral cell suspension were sorted into different cellular populations using fluorescent cell markers that are non-species specific. Over 30 different cell markers were tested. Additionally, cell suspension from Aiptasia pallida was also tested, and a phagocytosis test was done as a downstream functional assay.
We found that 24 of the screened markers positively labeled coral cells and 16 differentiated cell sub-populations. We identified 12 different cellular sub-populations using three markers, and found that each sub-population is primarily homogeneous. Lastly, we verified this technique in a sea anemone, Aiptasia pallida, and found that with minor modifications, a similar gating strategy can be successfully applied. Additionally, within A. pallida, we show elevated phagocytosis of sorted cells based on an immune associated marker.
In this study, we successfully adapted FACS for isolating coral cell populations and conclude that this technique is translatable for future use in other species. This technique has the potential to be used for different types of studies on the cellular stress response and other immunological studies.
目前，用于理解非模式物种细胞生物学的通用方法非常少见，但却十分必要。为了解决这一问题，我们对传统上用于生物医学领域的一项技术进行了改进，将其应用于生态和进化研究。荧光激活细胞分选术（FACS）常用于分选和鉴定细胞群体。在本研究中，我们开发了一种方法来鉴定和分离珊瑚及其他刺胞动物中的不同细胞群体。
利用荧光激活细胞分选术（FACS），使用非物种特异性的荧光细胞标记物将珊瑚细胞悬液分选到不同的细胞群体中。测试了30多种不同的细胞标记物。此外，还对苍白艾氏海葵的细胞悬液进行了测试，并进行了吞噬试验作为下游功能分析。
我们发现，筛选出的标记物中有24种能对珊瑚细胞进行阳性标记，可区分出16个细胞亚群。我们使用三种标记物鉴定出12个不同细胞亚群，并发现每个亚群基本都是同质的。最后，我们在海葵苍白艾氏海葵中验证了该技术，发现只需进行少量修改，就能成功应用类似的门控策略。此外，在苍白艾氏海葵中，基于一种免疫相关标记物，我们发现分选后的细胞吞噬作用增强。
在本研究中，我们成功地将FACS技术应用于分离珊瑚细胞群体，并得出该技术可应用于未来其他物种研究的结论。该技术有潜力用于细胞应激反应的不同类型研究及其他免疫学研究。
