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CD44 serving as marker of hematopoietic stem/progenitor cells and application thereof

A CD44, 1.CD44 technology, applied in the field of markers of hematopoietic stem/progenitor cells, can solve the problem of CD43 not being well identified

Pending Publication Date: 2021-10-26
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] CD43 differentiated in vitro + The cell population is heterogeneous, including various stages of hematopoietic cells produced by primitive hematopoiesis and definitive hematopoiesis, and only some of the cells produced by permanent hematopoiesis have the multi-lineage differentiation potential of hematopoietic stem / progenitor cells, so CD43 Cannot identify and enrich hematopoietic stem / progenitor cell populations differentiated in vitro

Method used

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  • CD44 serving as marker of hematopoietic stem/progenitor cells and application thereof
  • CD44 serving as marker of hematopoietic stem/progenitor cells and application thereof
  • CD44 serving as marker of hematopoietic stem/progenitor cells and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Using human pluripotent stem cells (hPSCs) for in vitro differentiation culture to obtain hematopoietic stem / progenitor cells, and verified CD44 + The cells have the multi-lineage differentiation potential of hematopoietic stem / progenitor cells. Specific steps are as follows:

[0046] (1) hPSCs were subjected to hematopoietic monolayer differentiation in vitro, and after the eighth day of differentiation, they were sorted by surface antibodies CD43 and CD44, and CD43 was sorted out + CD44 + and CD43 + CD44 - The two populations of cells were then subjected to subsequent colony formation experiments (CFU), in vitro co-culture differentiation experiments, and in vivo bone marrow cavity differentiation experiments to detect the differentiation potential of these two types of cells in each lineage of blood cells (such as figure 1 shown).

[0047] (2) CFU experiment is one of the effective methods to detect the proliferation and differentiation ability of cult...

Embodiment 2

[0068] Example 2 Isolate and obtain hematopoietic stem / progenitor cells from human peripheral blood, and combine CD34 and CD44 markers for characterization, and verify that CD34 + CD44 + Hematopoietic stem / progenitor cells have multi-lineage differentiation potential. Specifically include the following steps:

[0069] (1) if Figure 5 As shown, after the mobilization of human peripheral blood (that is, through drug treatment, the hematopoietic stem / progenitor cells in the bone marrow enter the peripheral blood, thereby increasing the content of hematopoietic stem / progenitor cells in the peripheral blood), the mononuclear cells obtained Then use CD34 antibody magnetic beads for primary purification, and then use surface antibodies CD34 and CD44 for sorting, and sort out CD34 + CD44 + and CD34 + CD44 - The two populations of cells were then subjected to subsequent colony formation experiments (CFU), in vitro co-culture differentiation experiments, and in vivo bone marrow c...

Embodiment 3

[0087] Human pluripotent stem cells (hPSCs) were used to differentiate in vitro to obtain hematopoietic stem / progenitor cells, and CD43 was verified + CD44 + The cells have the characteristics of hematopoietic stem / progenitor cells produced by permanent hematopoiesis.

[0088] Specific steps are as follows:

[0089] (1) The hPSCs were differentiated into monolayer hematopoietic in vitro, and after the eighth day of differentiation, the following three experiments were performed on the adherent cells: ① Immunofluorescence analysis of newborn HS / PC; ② Flow cytometric analysis of newborn HS / PC ③Use the surface antibodies CD31, CD43 and CD44 to sort, and sort out CD31 + CD43 - CD44 - ,CD31 + CD43 - CD44 + ,CD31 + CD43 + CD44 - and CD31 + CD43 + CD44 + Four populations of cells were then subjected to subsequent colony formation experiments (CFU), gene expression analysis, and in vitro culture experiments. The above experiments were designed to detect the ability of CD...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to CD44 serving as a marker of hematopoietic stem / progenitor cells and application thereof. The CD44 is used as the marker of the hematopoietic stem / progenitor cells, can be used for distinguishing original hematopoiesis from permanent hematopoiesis, and is used for separating and enriching the hematopoietic stem / progenitor cells from in-vivo sources and the hematopoietic stem / progenitor cells which are obtained by in-vitro differentiation and have multi-line differentiation potential.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to CD44 as a marker of hematopoietic stem / progenitor cells and its application. Background technique [0002] Hematopoietic stem / progenitor cell transplantation is an effective treatment for a variety of blood system diseases, but the hematopoietic stem / progenitor cells currently used for transplantation therapy face problems such as source limitation and immune rejection, and the use of human pluripotent stem cells through in vitro differentiation Methods Obtaining hematopoietic stem / progenitor cells for transplantation therapy can solve these problems well. In the current in vitro differentiation system, CD43 is used as the surface marker of hematopoietic stem / progenitor cells, and CD34 is used in adult bone marrow and peripheral blood. + The cell population serves as the population of hematopoietic stem / progenitor cells. [0003] CD43 differentiated in vitro + The cell...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68C07K16/28C12N5/0789A61K39/395A61P35/02A61P7/06A61P37/02
CPCG01N33/6872G01N33/6893G01N33/56966C07K16/2884C12N5/0647A61P35/02A61P7/06A61P37/02G01N2333/70585G01N2800/22C12N2509/00A61K2039/505
Inventor 潘光锦朱艳玲谷佳明王天宇黄可张田
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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