| Differentiation of Human Induced Pluripotent Stem Cells (hiPSCs) into Osteoclasts
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Author:
Date:
2020-12-20
[Abstract] Defects in bone resorption by osteoclasts result in numerous rare genetic bone disorders as well as in some common diseases such as osteoporosis or osteopetrosis. The use of hiPSC-differentiated osteoclasts opens new avenues in this research field by providing an unlimited cell source and overcoming obstacles such as unavailability of human specimens and suitable animal models. Generation of hiPSCs is well established but efficient differentiation of hiPSCs into osteoclasts has been challenging. Published hiPSC-osteoclast differentiation protocols use a hiPSC-OP9 co-culture system or hiPSC-derived embryoid bodies (EBs) with multiple cytokines. Our three-stage protocol consists of 1) EB mesoderm differentiation, 2) expansion of myelomonocytic cells and 3) maturation of hiPSC-osteoclasts. ...
[摘要] [摘要]破骨细胞引起的骨吸收缺陷导致许多罕见的遗传性骨疾病以及某些常见的疾病，例如骨质疏松症或骨质疏松症。采用的hiPSC -分化破骨细胞通过提供无限的细胞来源和克服障碍，如人体标本和合适的动物模型的可用性打开了该领域的新途径。hiPSC的生成已被公认，但是将hiPSC高效分化为破骨细胞一直具有挑战性。发布的hiPSC -osteoclast分化协议使用的hiPSC-OP9共培养体系或hiPSC细胞来源的胚状 具有多种细胞因子的机体（EB）。我们的三阶段协议包含：1）中胚层EB分化，2）的扩张骨髓单核细胞和3）的成熟的hiPSC -osteoclasts。我们通过在Nunclon Sphera微孔板上培养Accutase分离的hiPSCs来产生大小均一的EB，并在4天的细胞因子混合物中促进EB中胚层分化。对于第2阶段，将EBs转移至明胶包被的平板中，并用hM -CSF和hIL-3培养，以扩增骨髓单核细胞群。通过与维生素d，补充hTGF β，HM -CSF和hRANKL ，在第2阶段结束时收集的细胞的diff erentiated成成熟破骨细胞（第3阶段）。与其他技术相比，我们的协议不需要共培养系统。诱导EBs分化为中胚层 均匀的方式; 使用较少的细胞因子进行分化；只需要很短的时间就可以使破骨细胞成熟，并产生足够数量的破骨细胞用于后续的分子分析。
图形摘要： ...
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| Mouse ESC Differentiation to Nkx2.1+ Lung and Thyroid Progenitors
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Author:
Date:
2012-11-20
[Abstract] The de novo derivation of lung progenitors from pluripotent stem cells provides the opportunity to model early lung development in vitro and allows easy access to cells for tissue engineering or basic cell biology studies. This detailed protocol allows the generation of lung and thyroid progenitors from mouse embryonic stem cell (ESC) or induced pluripotent stem cell (iPSC) lines. When used together with a published Nkx2.1-GFP knock-in ESC line, the protocol allows tracking and purification of lung and thyroid progenitors by sorting on the GFP reporter based on the induction of the earliest known marker of lung and thyroid cell fate, Nkx2.1. After sorting, a pure population of Nkx2.1+ cells can then be replated for further expansion, differentiation, and maturation in ...
[摘要] 来自多能干细胞的肺祖细胞的新发现提供了在体外模拟早期肺发育的机会，并且容易获得用于组织工程或基础细胞生物学研究的细胞。 该详细方案允许从小鼠胚胎干细胞（ESC）或诱导多能干细胞（iPSC）系生成肺和甲状腺祖细胞。 当与公布的Nkx2.1-GFP敲入ESC系一起使用时，该方案允许通过基于对最早已知的肺和甲状腺细胞命运的标记物的诱导在GFP报告基因上进行分选来跟踪和纯化肺和甲状腺祖细胞， Nkx2.1。 在分选后，然后可以在无血清条件下培养纯化的Nkx2.1 +细胞群，以进一步扩增，分化和成熟。
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| Mouse Embryonic Stem Cell Differentiation to Hematopoietic Precursors
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Author:
Date:
2012-04-05
[Abstract] Embryonic stem cells are derived from inner cell mass of an embryo that can differentiate into every cell type in the body. Clinically, cultured red blood cell supply is of great interest. However, some of the hurdles need to be overcome. This protocol describes a two step protocol to form embryoid body and differentiate them in to hematopoietic lineage cells, especially erythroid cells.
[摘要] 胚胎干细胞是从小鼠囊胚期内细胞团分离得到。这些内细胞团可以分化成身体内每一种细胞类型。红细胞培养在临床上引起了极大兴趣。然而，有一些障碍需要克服。这份指南介绍了两部分内容，胚体形成和将其分化到造血系细胞，特别是红系细胞。
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