Directed differentiation of human pluripotent stem cells into epidermal stem and progenitor cells.
Directed differentiation of human pluripotent stem cells into epidermal stem and progenitor cells.
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DOI：
10.1007/s11033-021-06588-3
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发表时间：
2021-08
影响因子：
2.8
通讯作者：
Wells SI
中科院分区：
文献类型：
--
作者：
Ruiz-Torres S;Lambert PF;Wikenheiser-Brokamp KA;Wells SI
通过体细胞重编程产生的诱导多能干细胞（PSC）在培养中自我更新，原则上可以分化为任何细胞类型。这些细胞代表了疾病建模、药物筛选、再生医学和发现治疗组织特异性病理的个性化疗法的强大工具。我们以前曾报道过将人PSC定向分化为表皮干细胞和祖细胞（ESPC）和3D分层表皮，以模拟遗传性范可尼贫血综合征（FA）。在FA患者人群中验证了该已发表系统中发现的表皮细胞连接缺陷。在本文中，我们详细描述了使用不同的PSC系（209.2 PSC）产生PSC衍生的角质形成细胞的可再生来源的相应的20天方案。这种方法修改了以前的方案，以最大限度地减少自发性细胞死亡和终末分化，消除细胞应激诱导角质形成细胞选择步骤，并减少总方案持续时间和成本。通过添加相关形态发生剂和Rho相关蛋白激酶（ROCK）抑制剂，将独立供体来源的PSC系转化为ESPC。与先前发表的FA PSC系相比，正常209.2 PSC系的结果突出显示了2D中的表皮，并且还显示了所得3D表皮中的可变特征。209.2基于标记物分析，PSC衍生的ESPC表现出基底细胞表型，同时保持被工程化为具有与胎儿表皮一致的形态的PSC衍生的表皮器官型筏的能力。此外，确定角质形成细胞身份的转录分析表明，209.2 PSC衍生的ESPC表达表皮选择性标志物，而不是早期内胚层标志物，因此支持p63+表皮发育的不成熟阶段。该方案为人类ESPC和衍生3D表皮模型的生成提供了加速路径，以研究正常表皮发育和稳态，阐明表皮疾病发病机制，并为开发个性化治疗提供平台。
Induced pluripotent stem cells (PSCs) produced by somatic cell reprogramming self-renew in culture and can, in principle, differentiate into any cell type. These cells represent a powerful tool for disease modeling, drug screening, regenerative medicine, and the discovery of personalized therapies to treat tissue-specific pathologies. We have previously reported the directed differentiation of human PSCs into epidermal stem and progenitor cells (ESPCs) and 3D stratified epidermis to model the inherited syndrome Fanconi anemia (FA). Epidermal cell junctional defects discovered in this published system were validated in FA patient populations. Herein, we describe in detail the corresponding 20-day protocol for generating a renewable source of PSC-derived keratinocytes using a distinct PSC line (209.2 PSC). This approach modifies previous protocols to minimize spontaneous cell death and terminal differentiation, eliminate cell stress-inducing keratinocyte selection steps, and reduce total protocol duration and cost. Independent donor-derived PSC lines were converted into ESPCs through the addition of relevant morphogens and a Rho-associated protein kinase (ROCK) inhibitor. Results for the normal 209.2 PSC line highlight consistencies in 2D and also variable features in the resulting 3D epidermis compared to the previously published FA PSC lines. 209.2 PSC-derived ESPCs exhibited a basal cell phenotype based on marker analysis, while maintaining the capacity to be engineered into PSC-derived epidermal organotypic rafts with morphology consistent with fetal epidermis. Furthermore, transcriptional analyses to determine keratinocyte identity demonstrated 209.2 PSC-derived ESPCs express epidermis-selective markers and not early endoderm markers, thus supporting an immature stage of p63+ epidermal development. This protocol provides an accelerated path for the generation of human ESPCs and derivative 3D epidermal models to study normal epidermal development and homeostasis, elucidate mechanisms of epidermal disease pathogenesis, and provides a platform for developing personalized therapies.
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影响因子：
5.2
作者：
Dabelsteen, Sally;Hercule, Paula;Barron, Patricia;Rice, Meghan;Dorsainville, Gregory;Rheinwald, James G.
通讯作者：
Rheinwald, James G.
影响因子：
2.4
作者：
Fisher JB;Pulakanti K;Rao S;Duncan SA
通讯作者：
Duncan SA
影响因子：
5.8
作者：
Schmook, FP;Meingassner, JG;Billich, A
通讯作者：
Billich, A
影响因子：
9.2
作者：
McMullan, R;Lax, S;Hotchin, NA
通讯作者：
Hotchin, NA
DOI：
10.1387/ijdb.170341fw
发表时间：
2018
期刊：
The International journal of developmental biology
影响因子：
--
作者：
Belokhvostova D;Berzanskyte I;Cujba AM;Jowett G;Marshall L;Prueller J;Watt FM
通讯作者：
Watt FM