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在自组装肽水凝胶的 3D 培养系统中从小鼠多能干细胞生成造血细胞。

Generation of hematopoietic cells from mouse pluripotent stem cells in a 3D culture system of self-assembling peptide hydrogel.

机构信息

Bone Marrow Transplantation Center, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang, China.

Key Laboratory of Regenerative Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.

出版信息

J Cell Physiol. 2020 Mar;235(3):2080-2090. doi: 10.1002/jcp.29110. Epub 2019 Aug 6.

Abstract

In vitro generation of hematopoietic stem cells from pluripotent stem cells (PSCs) can be regarded as novel therapeutic approaches for replacing bone marrow transplantation without immune rejection or graft versus host disease. To date, many different approaches have been evaluated in terms of directing PSCs toward different hematopoietic cell types, yet, low efficiency and no function restrict the further hematopoietic differentiation study, our research aims to develop a three dimension (3D) hematopoietic differentiation approach that serves as recapitulation of embryonic development in vitro to a degree of complexity not achievable in a two dimension culture system. We first found that mouse PSCs could be efficiently induced to hematopoietic differentiation with an expression of hematopoietic makers, such as c-kit, CD41, and CD45 within self-assembling peptide hydrogel. Colony-forming cells assay results suggested mouse PSCs (mPSCs) could be differentiated into multipotential progenitor cells and 3D induction system derived hematopoietic colonies owned potential of differentiating into lymphocyte cells. In addition, in vivo animal transplantation experiment showed that mPSCs (CD45.2) could be embedded into nonobese diabetic/severe combined immunodeficiency mice (CD45.1) with about 3% engraftment efficiency after 3 weeks transplantation. This study demonstrated that we developed the 3D induction approach that could efficiently promote the hematopoietic differentiation of mPSCs in vitro and obtained the multipotential progenitors that possessed the short-term engraftment potential.

摘要

体外诱导多能干细胞(PSCs)生成造血干细胞可以被视为替代骨髓移植的新治疗方法,不会发生免疫排斥或移植物抗宿主病。迄今为止,已经评估了许多不同的方法来指导 PSCs 向不同的造血细胞类型分化,但效率低和无功能限制了进一步的造血分化研究,我们的研究旨在开发一种三维(3D)造血分化方法,该方法在一定程度上可以模拟体外胚胎发育,而二维培养系统则无法实现如此复杂的程度。我们首先发现,在自组装肽水凝胶中,小鼠 PSCs 可以有效地被诱导分化为造血细胞,并表达造血标志物,如 c-kit、CD41 和 CD45。集落形成细胞检测结果表明,小鼠 PSCs(mPSCs)可以分化为多能祖细胞,并且 3D 诱导系统衍生的造血集落具有分化为淋巴细胞的潜力。此外,体内动物移植实验表明,在 3 周的移植后,mPSCs(CD45.2)可以被嵌入非肥胖型糖尿病/严重联合免疫缺陷小鼠(CD45.1)中,植入效率约为 3%。这项研究表明,我们开发了一种 3D 诱导方法,可以有效地促进 mPSCs 在体外的造血分化,并获得了具有短期植入潜力的多能祖细胞。

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