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Method for separating and enriching hematopoietic stem cells in peripheral blood

A technology for separating and enriching hematopoietic stem cells, which is applied to the field of separating and enriching hematopoietic stem cells from peripheral blood that is not mobilized by bone marrow, can solve the problems of bone marrow sampling trauma, unfavorable expansion and culture, and the number and purity of hematopoietic stem cells are not high, so as to solve the problem of obtaining materials. Inconvenient, pain-avoiding effects

Inactive Publication Date: 2016-03-23
王泰华
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Problems solved by technology

[0006] That is to say, among the three methods, bone marrow extraction will cause trauma, and patients need to endure great pain; extraction from mobilized peripheral blood requires the use of mobilizing agents, and there will be related side effects; there are some aspects of control over the source of umbilical cord blood.
Therefore, there are certain defects in the three kinds of materials
Moreover, the content of hematopoietic stem cells in non-mobilized peripheral blood is small, so the quantity and purity of hematopoietic stem cells extracted from non-mobilized peripheral blood are not high, which is not conducive to later in vitro expansion and culture

Method used

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  • Method for separating and enriching hematopoietic stem cells in peripheral blood
  • Method for separating and enriching hematopoietic stem cells in peripheral blood
  • Method for separating and enriching hematopoietic stem cells in peripheral blood

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Embodiment Construction

[0052] The invention extracts HSCs from non-mobilized peripheral blood, avoids the problems in the prior art, and opens up a new source.

[0053] In recent years, the development of HSC surface marker research has enriched the means of stem cell isolation and purification. According to the surface markers of HSC, hematopoietic stem cells can be detected by various cell biology and molecular biology methods such as immunocytochemistry, flow cytometry, molecular hybridization and PCR. As mentioned above, most of the screening of hematopoietic stem cells is still based on the enrichment of CD34+ cells. For example, the purity of fluorescence activated cell sorting (fluorescence activated cell sorting, FACS) can reach 95%, but the cost is higher and the instrument is also more expensive. Therefore, the present invention uses the immunoadsorption system--immunomagnetic beads to separate CD34+ cells, and the immunomagnetic bead separation system separates CD34+ cells. Large amount,...

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Abstract

The invention discloses a method for separating and enriching hematopoietic stem cells in peripheral blood. The method includes the first step of the autologous plasma preparation stage, the second step of the hydroxyethyl starch (HES) erythrocyte removal stage, the third step of mononuclear cell preparation, the fourth step of the mononuclear cell (MNC) in-vitro enrichment culture stage and the fifth step of the CD34+hematopoietic stem cell obtaining stage. Hematopoietic stem cells large in number and high in purity are separated in vitro from materials easily and conveniently taken, and therefore later-period large-amount amplification culture is facilitated, and the method can be used for hematopoietic stem cell transplantation to treat clinical hematopoietic system diseases.

Description

technical field [0001] The invention belongs to the technical field of stem cells, and in particular relates to a method for separating and enriching hematopoietic stem cells in peripheral blood, specifically also belongs to the field of biotechnology, in particular to a method for separating and enriching hematopoietic stem cells in non-bone marrow mobilized peripheral blood. Background technique [0002] At present, most biologists and medical scientists believe that stem cells are a type of cells that come from embryos, fetuses or adults with unlimited self-renewal, proliferation and differentiation capabilities under certain conditions, and can produce phenotype and genotype and their own complete The same daughter cells can also produce specialized cells that make up body tissues and organs, and can also differentiate into progenitor cells. [0003] Hematopoietic stem cells (HSCs) are hematopoietic precursor cells with high self-renewal ability and multi-directional dif...

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Application Information

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IPC IPC(8): C12N5/0789
Inventor 王泰华
Owner 王泰华
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