Preparation method for hematopoietic progenitor cells and special medium for same

A technology for hematopoietic progenitor cells and cell culture, which is applied to the preparation method of hematopoietic progenitor cells and the field of special culture medium thereof, can solve the problems of indeterminate differentiation direction and low differentiation efficiency, and achieve the effect of clear components and promotion of efficient production.

Inactive Publication Date: 2012-10-17
PEKING UNIV +1
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  • Claims
  • Application Information

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Problems solved by technology

Existing studies have reported methods for the differentiation of human pluripotent stem cells in vitro to produce hematopoietic progenitor cells. They mainly adopt co-cultivation with stromal cells and differentiation of embryonic bodies. These differentiation methods have many disadvantages, such as differentiation The direction is not oriented, the differentiation efficiency is low, and the differentiation conditions contain serum and mouse stromal cells, etc., which will seriously limit the application of hematopoietic cells produced by this differentiation method in the treatment of blood diseases

Method used

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  • Preparation method for hematopoietic progenitor cells and special medium for same
  • Preparation method for hematopoietic progenitor cells and special medium for same
  • Preparation method for hematopoietic progenitor cells and special medium for same

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Embodiment 1

[0047] Example 1. Exploration of system conditions for directed differentiation of human embryonic stem cells to hematopoietic progenitor cells

[0048] 1. Establishment of differentiation system

[0049] By tracking the occurrence of hematopoietic stem cells in vivo, it was proved that hematopoietic endothelial cells can directly generate hematopoietic stem cells.

[0050] According to this discovery, using the in vitro differentiation system of human embryonic stem cells, it was found that mesoderm cells (KDR+) differentiated from human embryonic stem cells will further differentiate to produce a group of cells with endothelial cell properties and hematopoietic cells, and this group of cells is named hematopoietic endothelial cells . Hematopoietic endothelial cells are mainly enriched in the CD31+ cell population, which can directly produce CD45+ hematopoietic cells, and express CD34+, KDR+, Tie-2, Ac-LDL and PDGFa- and other surface markers ( figure 1 The differentiation ...

Embodiment 2

[0072] Example 2, Directed differentiation and identification of human embryonic stem cells to hematopoietic progenitor cells

[0073] Human embryonic stem cell line H1 was purchased from Wicell Corporation, USA.

[0074] (1) Directed differentiation of human embryonic stem cells to hematopoietic progenitor cells (the pH value of the cell culture medium is between 7.0-7.2)

[0075] 1. Method 1:

[0076] After studying the generation signal of blood endothelial cells and the signal regulating the differentiation of hematopoietic endothelial cells to new hematopoietic progenitor cells, according to the above-mentioned new step-by-step differentiation method, the directional differentiation of human embryonic stem cells to hematopoietic progenitor cells is completed, as follows:

[0077] 1) Make the growth density up to (1X10e 4 ) human embryonic stem cells H1 were inoculated onto Matrigel (BD, catalog number 356230) treated 12-well plate (BD Falcon), and then added human embry...

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Abstract

The invention discloses a preparation method for hematopoietic progenitor cells and a special medium for the same. The invention provides the medium for hematopoietic progenitor cells prepared through differentiation of human embryonic stem cells or induced pluripotent stem cells, and the medium comprises cell culture fluid I, cell culture fluid II, cell culture fluid III and cell culture fluid IV. According to results of experiments in the invention, a novel system which has definite components and induces differentiation of human embryonic stem cells step by step to generate hematopoietic progenitor cells is established; the system provides a good research platform for further research of differentiation and generation of hematopoietic stem / progenitor cells and provides a differentiation method for obtaining hematopoietic progenitor cells which can be used in treatment of clinical blood diseases since the differentiation system utilizes serum-free and murine-free stromal cells and the like.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a preparation method of hematopoietic progenitor cells and a special culture medium thereof. Background technique [0002] Human pluripotent stem cells (human embryonic stem cells and human induced pluripotent stem cells) have the ability to proliferate indefinitely in vitro, and can differentiate to produce various cell types such as blood cells, which will provide a new source of cells for the treatment of blood diseases. Existing studies have reported methods for the differentiation of human pluripotent stem cells in vitro to produce hematopoietic progenitor cells. They mainly adopt co-cultivation with stromal cells and differentiation of embryonic bodies. These differentiation methods have many disadvantages, such as differentiation The direction is not oriented, the differentiation efficiency is low, and the differentiation conditions contain serum and mouse-derived stromal cell...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0789
Inventor 邓宏魁王承艳汤旭明苗振川孙晓萌吕娅歆尹明
Owner PEKING UNIV
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