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Method for preparing macrophage by iPS cell differentiation

A macrophage and cell technology, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve problems such as lack of function, inability to self-renew, and difficulty in obtaining

Inactive Publication Date: 2019-07-05
杭州荣泽生物科技集团有限公司
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Problems solved by technology

The former can proliferate indefinitely, but the source of the tumor may lead to changes in genetic structure, resulting in loss of function; the latter cannot self-renew, and each test requires a large amount of peripheral blood, which is difficult to obtain and the quantity is small, and different sources and batches will also cause results non-repeatability

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  • Method for preparing macrophage by iPS cell differentiation
  • Method for preparing macrophage by iPS cell differentiation
  • Method for preparing macrophage by iPS cell differentiation

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Embodiment Construction

[0050] The invention provides a method for obtaining a large amount of macrophages with immune function from iPS cells. Specifically, the invention provides a method system for suspending culture of iPS into embryoid body EBs, and then differentiating them into macrophages through the EB-induced pathway, which specifically includes cell culture of hiPS, identification of hiPS, and identification of hiPS to form EBs. Methods, culture and identification, induction of hiPS-derived EBs to differentiate into macrophages, and identification of hiPS-derived macrophages.

[0051] hiPS cell culture

[0052] Dissolve 100 μL Matrigel in 10 mL DMEM basal medium, add 2 mL to each 35 mm cell culture dish, place at room temperature for 2 hours, and store at 4°C. hiPS (DYR0100, Stem Cell Bank, Chinese Academy of Sciences) were cultured in hiPS feeder-free complete medium (Pluripotent Stem CellSFM XF / FF, Stem Cell Bank, Chinese Academy of Sciences) with 10 μM Y27632 (ROCK kinase inhibitor, ...

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Abstract

The invention aims to provide a method for preparing mature macrophages by effectively differentiating and inducing iPS cells in vitro. The invention provides a method system for suspending and culturing iPS into embryoid-like EBs and differentiating the embryoid-like EBs into macrophages through an EB induction way so as to obtain a large number of macrophages with an immune function from iPS. Inaddition, the method for inducing differentiation of EB is provided to further culture under conditions suitable for induction of differentiation of blood cells, and is capable of producing various blood cells.

Description

[0001] 【Technical field】 [0002] The invention relates to a method for preparing macrophages by differentiation of iPS (induced pluripotent stem cells). [0003] 【Background technique】 [0004] Human macrophages (macrophage, ) research is the limited and heterogeneous sources of human macrophages. At present, human macrophages used in experimental research are mostly derived from tumor cell lines and peripheral blood mononuclear cells. The former can proliferate indefinitely, but the source of the tumor may lead to changes in genetic structure, resulting in loss of function; the latter cannot self-renew, and each test requires a large amount of peripheral blood, which is difficult to obtain and the quantity is small, and different sources and batches will also cause results non-repeatability. How to obtain a large number of functionally active macrophages is the key to research, and the emergence of induced pluripotent stem cells (iPS) technology provides a new idea for obt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0786
CPCC12N5/0645C12N2501/22C12N2501/2303C12N2506/45
Inventor 陈相波雷鸣田朋飞
Owner 杭州荣泽生物科技集团有限公司
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