Coral cell separation and isolation by fluorescence-activated cell sorting (FACS).
Coral cell separation and isolation by fluorescence-activated cell sorting (FACS).
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通过荧光激活的细胞分选（FACS）分离珊瑚细胞分离和分离。
DOI：
10.1186/s12860-017-0146-8
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发表时间：
2017-08-29
期刊：
影响因子：
--
通讯作者：
Traylor-Knowles N
中科院分区：
文献类型：
--
作者：
Rosental B;Kozhekbaeva Z;Fernhoff N;Tsai JM;Traylor-Knowles N
理解非模式物种细胞生物学的通用方法非常罕见，但非常需要。为了解决这个问题，我们修改了一种传统上用于生物医学领域的生态和进化研究的技术。荧光激活细胞分选(FACS)通常用于细胞群体的分选和鉴定。在这项研究中，我们开发了一种方法来鉴定和分离珊瑚和其他线虫的不同细胞群。通过荧光激活细胞分选(FACS)，珊瑚细胞悬浮液通过非物种特异性的荧光细胞标记被分选到不同的细胞群中。测试了30多种不同的细胞标志物。此外，还对苍白拟青霉的细胞悬浮液进行了测试，并进行了吞噬实验作为下游功能检测。我们发现，在筛选的DNA标记中，有24个珊瑚细胞和16个分化细胞亚群呈阳性标记。我们利用三个标记对12个不同的细胞亚群进行了鉴定，发现每个亚群基本上是同质的。最后，我们在海葵Aiptasia and pallida上验证了这项技术，发现只需稍加修改，就可以成功地应用类似的门控策略。此外，在梅毒螺旋体内，我们发现基于免疫相关标记物的分选细胞吞噬功能增强。在这项研究中，我们成功地将FACS用于分离珊瑚细胞种群，并得出结论，该技术可移植到未来用于其他物种。这项技术有可能用于不同类型的细胞应激反应研究和其他免疫学研究。本文的在线版本(10.1186/s12860-0170146-8)包含补充材料，可供授权用户使用。
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. The online version of this article (10.1186/s12860-017-0146-8) contains supplementary material, which is available to authorized users.
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影响因子：
3.7
作者：
Alieva NO;Konzen KA;Field SF;Meleshkevitch EA;Hunt ME;Beltran-Ramirez V;Miller DJ;Wiedenmann J;Salih A;Matz MV
通讯作者：
Matz MV
影响因子：
32.4
作者：
Garg S;Sharma M;Ung C;Tuli A;Barral DC;Hava DL;Veerapen N;Besra GS;Hacohen N;Brenner MB
通讯作者：
Brenner MB
DOI：
10.1073/pnas.211439698
发表时间：
2001-10-09
影响因子：
11.1
作者：
Domart-Coulon, IJ;Elbert, DC;Ostrander, GK
通讯作者：
Ostrander, GK
影响因子：
9.2
作者：
Hughes, Terence P.;Rodrigues, Maria J.;Willis, Bette
通讯作者：
Willis, Bette
影响因子：
3
作者：
Bertucci, Anthony;Tambutte, Sylvie;Zoccola, Didier
通讯作者：
Zoccola, Didier