Isolation of Human Photoreceptor Precursors via a Cell Surface Marker Panel from Stem Cell-Derived Retinal Organoids and Fetal Retinae.
Isolation of Human Photoreceptor Precursors via a Cell Surface Marker Panel from Stem Cell-Derived Retinal Organoids and Fetal Retinae.
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DOI：
10.1002/stem.2775
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发表时间：
2018-05
期刊：
影响因子：
--
通讯作者：
Sowden JC
中科院分区：
文献类型：
--
作者：
Lakowski J;Welby E;Budinger D;Di Marco F;Di Foggia V;Bainbridge JWB;Wallace K;Gamm DM;Ali RR;Sowden JC
在发达国家，由于视网膜变性导致的感光细胞损失是失明的主要原因之一。虽然目前没有有效的治疗方法，但使用干细胞衍生的感光细胞的细胞替代疗法可能是未来可行的治疗选择。为了确保这种方法的安全性和有效性，必须开发稳健的细胞分离和纯化方案。为此，我们先前开发了一种生物标志物面板，用于从发育中的小鼠视网膜和小鼠胚胎干细胞培养物中分离小鼠光感受器前体。在目前的研究中，我们将这种方法应用于人类多能干细胞（hPSC）系统，并确定了可用于分离人类光感受器的新型生物标志物组合。使用高通量流式细胞术方法针对242种人单克隆抗体筛选人视网膜样品和hPSC衍生的视网膜类器官培养物。我们鉴定了在人视网膜和hPSC分化培养物中具有显著表达水平的46种生物标志物。使用显示在离散细胞群中表达的所选候选生物标志物，对来自胎儿组织或源自胚胎和诱导多能干细胞培养物的人视网膜细胞样品进行荧光激活细胞分选（FACS）。通过使用感光细胞特异性抗体和Ki-67的免疫细胞化学分析证明了感光细胞的富集和有丝分裂活性细胞的排除。我们建立了一种生物标志物组合，其能够从人视网膜和hPSC衍生的类器官培养物中稳健地纯化活的人光感受器。干细胞2018;36：709-722
Loss of photoreceptor cells due to retinal degeneration is one of the main causes of blindness in the developed world. Although there is currently no effective treatment, cell replacement therapy using stem‐cell‐derived photoreceptor cells may be a feasible future treatment option. In order to ensure safety and efficacy of this approach, robust cell isolation and purification protocols must be developed. To this end, we previously developed a biomarker panel for the isolation of mouse photoreceptor precursors from the developing mouse retina and mouse embryonic stem cell cultures. In the current study we applied this approach to the human pluripotent stem cell (hPSC) system, and identified novel biomarker combinations that can be leveraged for the isolation of human photoreceptors. Human retinal samples and hPSC‐derived retinal organoid cultures were screened against 242 human monoclonal antibodies using a high through‐put flow cytometry approach. We identified 46 biomarkers with significant expression levels in the human retina and hPSC differentiation cultures. Human retinal cell samples, either from fetal tissue or derived from embryonic and induced pluripotent stem cell cultures, were fluorescence‐activated cell sorted (FACS) using selected candidate biomarkers that showed expression in discrete cell populations. Enrichment for photoreceptors and exclusion of mitotically active cells was demonstrated by immunocytochemical analysis with photoreceptor‐specific antibodies and Ki‐67. We established a biomarker combination, which enables the robust purification of viable human photoreceptors from both human retinae and hPSC‐derived organoid cultures. Stem Cells 2018;36:709–722
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影响因子：
4.4
作者：
Klassen, HJ;Ng, TF;Young, MJ
通讯作者：
Young, MJ
影响因子：
64.8
作者：
Pearson, R. A.;Barber, A. C.;Rizzi, M.;Hippert, C.;Xue, T.;West, E. L.;Duran, Y.;Smith, A. J.;Chuang, J. Z.;Azam, S. A.;Luhmann, U. F. O.;Benucci, A.;Sung, C. H.;Bainbridge, J. W.;Carandini, M.;Yau, K. -W.;Sowden, J. C.;Ali, R. R.
通讯作者：
Ali, R. R.
影响因子：
3.5
作者：
Esumi N;Kachi S;Hackler L Jr;Masuda T;Yang Z;Campochiaro PA;Zack DJ
通讯作者：
Zack DJ
DOI：
10.1002/stem.2051
发表时间：
2015-08
期刊：
Stem cells (Dayton, Ohio)
影响因子：
--
作者：
Lakowski J;Gonzalez-Cordero A;West EL;Han YT;Welby E;Naeem A;Blackford SJ;Bainbridge JW;Pearson RA;Ali RR;Sowden JC
通讯作者：
Sowden JC
DOI：
10.1073/pnas.0905245106
发表时间：
2009-09-29
影响因子：
11.1
作者：
Meyer, Jason S.;Shearer, Rebecca L.;Gamm, David M.
通讯作者：
Gamm, David M.