Enhanced HSC-like cell generation from mouse pluripotent stem cells in a 3D induction system cocultured with stromal cells.
Enhanced HSC-like cell generation from mouse pluripotent stem cells in a 3D induction system cocultured with stromal cells.
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在与基质细胞共培养的 3D 诱导系统中增强小鼠多能干细胞的 HSC 样细胞生成
DOI：
10.1186/s13287-021-02434-2
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发表时间：
2021-06-19
影响因子：
7.5
通讯作者：
Huang H
中科院分区：
文献类型：
--
作者：
Shan W;Yu Q;Long Y;Luo Q;Li H;Han Y;Xu Y;Fu S;Zeng X;Wei C;Gao Y;Li X;Li X;Zhang L;Liu L;Chen M;Qian P;Huang H
数十年来，人们一直努力将多能干细胞（PSCs）分化为真正具有功能的造血干细胞（HSCs），然而，体外分化效率低以及体内造血重建效果不佳等问题依然存在，这主要归因于缺乏可靠、可重复或理想的分化体系。
在本研究中，我们通过三维诱导体系，随后与OP9基质细胞共培养，建立了一种体外分化体系，可使小鼠PSCs分化为具有体内造血重建能力的造血细胞。通过m - NSG移植试验评估源自小鼠PSCs的c - kit⁺细胞的体内造血重建潜力。采用流式细胞术分析、RNA测序和细胞周期分析来检测我们诱导体系中产生的内皮蛋白C受体（EPCR，CD201）细胞的体外造血能力。
经三维自组装肽诱导体系处理后再进行OP9共培养体系培养得到的c - kit⁺细胞，其体内再植活性明显优于经三维诱导体系处理后在0.1%明胶中培养得到的c - kit⁺细胞。我们有趣地发现，与CD201⁻c - kit⁺细胞相比，我们的3D + OP9体系富集了更高比例的CD201⁺c - kit⁺细胞，这些细胞在转录组水平和集落形成单位（CFU）形成能力等方面表现出更类似造血干细胞的特征，这在小鼠PSCs造血分化领域尚未见报道。此外，CD201⁺造血细胞处于相对缓慢的细胞周期状态，这与P57和Ccng2的高表达水平一致。进一步地，我们创新性地证明，Notch信号通路负责从小鼠PSCs体外诱导产生CD201⁺造血细胞。
总之，我们的研究结果为提高造血分化效率以及从小鼠PSCs生成用于临床应用的具有体内功能的类造血干细胞奠定了基础。
网络版本包含补充材料，获取网址：10.1186/s13287 - 021 - 02434 - 2 。
BackgroundDecades of efforts have attempted to differentiate the pluripotent stem cells (PSCs) into truly functional hematopoietic stem cells (HSCs), yet the problems of low differentiation efficiency in vitro and poor hematopoiesis reconstitution in vivo still exist, mainly attributing to the lack of solid, reproduced, or pursued differentiation system.MethodsIn this study, we established an in vitro differentiation system yielding in vivo hematopoietic reconstitution hematopoietic cells from mouse PSCs through a 3D induction system followed by coculture with OP9 stromal cells. The in vivo hematopoietic reconstitution potential of c-kit+cells derived from the mouse PSCs was evaluated via m-NSG transplantation assay. Flow cytometry analysis, RNA-seq, and cell cycle analysis were used to detect the in vitro hematopoietic ability of endothelial protein C receptor (EPCR, CD201) cells generated in our induction system.ResultsThe c-kit+cells from 3D self-assembling peptide induction system followed by the OP9 coculture system possessed apparently superiority in terms of in vivo repopulating activity than that of 3D induction system followed by the 0.1% gelatin culture. We interestingly found that our 3D+OP9 system enriched a higher percentage of CD201+c-kit+cells that showed more similar HSC-like features such as transcriptome level and CFU formation ability than CD201-c-kit+cells, which have not been reported in the field of mouse PSCs hematopoietic differentiation. Moreover, CD201+hematopoietic cells remained in a relatively slow cycling state, consistent with high expression levels of P57 and Ccng2. Further, we innovatively demonstrated that notch signaling pathway is responsible for in vitro CD201+hematopoietic cell induction from mouse PSCs.ConclusionsAltogether, our findings lay a foundation for improving the efficiency of hematopoietic differentiation and generating in vivo functional HSC-like cells from mouse PSCs for clinical application.
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影响因子：
64.5
作者：
Riddell J;Gazit R;Garrison BS;Guo G;Saadatpour A;Mandal PK;Ebina W;Volchkov P;Yuan GC;Orkin SH;Rossi DJ
通讯作者：
Rossi DJ
影响因子：
8.8
作者：
Park MA;Kumar A;Jung HS;Uenishi G;Moskvin OV;Thomson JA;Slukvin II
通讯作者：
Slukvin II
AGM地区发展中HSC谱系的分层组织和早期造血规范。
DOI：
10.1084/jem.20102419
发表时间：
2011-06-06
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Rybtsov S;Sobiesiak M;Taoudi S;Souilhol C;Senserrich J;Liakhovitskaia A;Ivanovs A;Frampton J;Zhao S;Medvinsky A
通讯作者：
Medvinsky A
影响因子：
5.9
作者：
Pearson, Stella;Cuvertino, Sara;Fleury, Maud;Lacaud, Georges;Kouskoff, Valerie
通讯作者：
Kouskoff, Valerie
影响因子：
64.8
作者：
Chen JY;Miyanishi M;Wang SK;Yamazaki S;Sinha R;Kao KS;Seita J;Sahoo D;Nakauchi H;Weissman IL
通讯作者：
Weissman IL