FLI1 and PKC co-activation promote highly efficient differentiation of human embryonic stem cells into endothelial-like cells.
FLI1 and PKC co-activation promote highly efficient differentiation of human embryonic stem cells into endothelial-like cells.
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FLI1和PKC共激活促进人胚胎干细胞高效分化为内皮样细胞
DOI：
10.1038/s41419-017-0162-9
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发表时间：
2018-01-26
影响因子：
9
通讯作者：
Hu L
中科院分区：
文献类型：
--
作者：
Zhao H;Zhao Y;Li Z;Ouyang Q;Sun Y;Zhou D;Xie P;Zeng S;Dong L;Wen H;Lu G;Lin G;Hu L
血管内皮细胞（EC）在各种再生过程中起重要作用，并且可用于各种治疗应用，例如心脏再生、基因治疗、组织工程化血管移植物和预血管化组织移植物。EC可以从多能干细胞和成体干细胞获得。以快速、高效、经济的方式从人胚胎干细胞（hESC）中获得EC。我们建立了一个条件性过表达系统的基础上，15个转录因子的报告是负责造血谱系。其中，只有FLI 1的过表达才能诱导hESC向造血细胞系分化。此外，同时过表达FLI 1和激活PKC在3天内快速有效地诱导hESC分化为诱导内皮细胞（iEC），而单独过表达FLI 1和激活PKC都不能从hESC衍生iEC。在诱导过程中，hESC分化成与EC外观一致的梭形细胞。流式细胞术分析显示，这些细胞中分别有92.2-98.9%和87.2-92.6%为CD 31+和CD 144+。在诱导过程中，血管特异性基因的表达显著增加，而多能性基因的表达逐渐减少。iEC掺入乙酰化低密度脂蛋白，强烈表达vWF并结合UEA-1。iEC在体外和体内也形成毛细血管样结构。RNA-seq分析证实这些细胞与其体内对应物非常相似。我们的研究结果表明，FLI 1和PKC的共激活可以诱导hESCs分化为iECs在快速，有效和经济的方式。
Rationale-endothelial cells (ECs) play important roles in various regeneration processes and can be used in a variety of therapeutic applications, such as cardiac regeneration, gene therapy, tissue-engineered vascular grafts and prevascularized tissue transplants. ECs can be acquired from pluripotent and adult stem cells. To acquire ECs from human embryonic stem cells (hESCs) in a fast, efficient and economic manner. We established a conditional overexpression system in hESCs based on 15 transcription factors reported to be responsible for hematopoiesis lineage. Among them, only overexpression of FLI1 could induce hESCs to a hematopoietic lineage. Moreover, simultaneous overexpression of FLI1 and activation of PKC rapidly and efficiently induced differentiation of hESCs into induced endothelial cells (iECs) within 3 days, while neither FLI1 overexpression nor PKC activation alone could derive iECs from hESCs. During induction, hESCs differentiated into spindle-like cells that were consistent in appearance with ECs. Flow cytometric analysis revealed that 92.2–98.9% and 87.2–92.6% of these cells were CD31+ and CD144+, respectively. Expression of vascular-specific genes dramatically increased, while the expression of pluripotency genes gradually decreased during induction. iECs incorporated acetylated low-density lipoproteins, strongly expressed vWF and bound UEA-1. iECs also formed capillary-like structures both in vitro and in vivo. RNA-seq analysis verified that these cells closely resembled their in vivo counterparts. Our results showed that co-activation of FLI1 and PKC could induce differentiation of hESCs into iECs in a fast, efficient and economic manner.
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影响因子：
64.5
作者：
Ginsberg M;James D;Ding BS;Nolan D;Geng F;Butler JM;Schachterle W;Pulijaal VR;Mathew S;Chasen ST;Xiang J;Rosenwaks Z;Shido K;Elemento O;Rabbany SY;Rafii S
通讯作者：
Rafii S
DOI：
10.1007/978-1-4939-6756-8_7
发表时间：
2017-01-01
期刊：
ADULT STEM CELLS: METHODS AND PROTOCOLS
影响因子：
--
作者：
Garikipati, Venkata N. S.;Kishore, Raj
通讯作者：
Kishore, Raj
DOI：
10.1161/atvbaha.112.301167
发表时间：
2013-06
期刊：
Arteriosclerosis, thrombosis, and vascular biology
影响因子：
--
作者：
Li J;Huang NF;Zou J;Laurent TJ;Lee JC;Okogbaa J;Cooke JP;Ding S
通讯作者：
Ding S
影响因子：
23.9
作者：
Pereira, Carlos-Filipe;Chang, Betty;Qiu, Jiajing;Niu, Xiaohong;Papatsenko, Dmitri;Hendry, Caroline E.;Clark, Neil R.;Nomura-Kitabayashi, Aya;Kovacic, Jason C.;Ma'ayan, Avi;Schaniel, Christoph;Lemischka, Ihor R.;Moore, Kateri
通讯作者：
Moore, Kateri
影响因子：
3.6
作者：
Orbay, Hakan;Devi, Kamaljit;Sahar, David E.
通讯作者：
Sahar, David E.