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Application of chrysin in in-vitro expansion of human hematopoietic stem cells

An in vitro amplification and artificial blood technology, applied in the field of cell culture, can solve problems such as poor quality, difficult hematopoietic reconstruction, and low success rate of engraftment, and achieve the effect of self-renewal promotion and self-renewal

Inactive Publication Date: 2020-01-10
INST OF HEMATOLOGY & BLOOD DISEASES HOSPITAL CHINESE ACADEMY OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the proportion of HSCs in umbilical cord blood only accounts for about 0.1% to 1%. Due to many thorny problems such as insufficient number, poor quality, and low success rate of engraftment of hematopoietic stem and progenitor cells, it is difficult to carry out effective transplantation of HSCs in patients after umbilical cord blood transplantation. Hematopoietic reconstruction largely restricts the development and clinical application of the stem cell industry
Previous studies have attempted to expand HSCs in vitro by adding a variety of cytokine combinations to the liquid medium, co-culturing with stromal cells, or culturing in a bioreactor, but the expansion effect of the above methods is still not ideal

Method used

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  • Application of chrysin in in-vitro expansion of human hematopoietic stem cells
  • Application of chrysin in in-vitro expansion of human hematopoietic stem cells
  • Application of chrysin in in-vitro expansion of human hematopoietic stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A kind of small molecular compound chrysin (chrysin C2968), its structure is:

[0029]

Embodiment 2

[0031] A method for in vitro expansion of human hematopoietic stem cells, comprising the steps of:

[0032] Freshly isolated human cord blood CD34 + Cells were resuspended in culture medium (Gibco Iscove's Modified Dulbecco's Medium+10% FBS+100ng / mL SCF+100ng / mL TPO+100ng / mL Flt3L+1%P / S) to a cell concentration of 5×10 4 Cells / mL, then spread in a 96-well plate, each well is 190 μL of cell suspension, and then add 10 μL of chrysin (C2968) small molecule compound diluted with the same culture medium, chrysin (C2968) small molecule compound The final concentration was 2.5 μM. at 37°C, 5% CO 2 Cultured in a constant temperature incubator for 7 days.

Embodiment 3

[0033] Example 3: Cell Phenotype Analysis

[0034] Using flow cytometry to detect the proportion and absolute number of CD34+CD38-, CD34+CD38-CD49f+, CD34+CD38-CD45RA-CD90+ cells to verify the effect of chrysin (C2968) small molecule compound on the expansion of hematopoietic stem cells in vitro, the results showed that poplar The C2968 small molecular compound has a significant effect on the expansion of hematopoietic stem cells in vitro.

[0035] Experimental method for detecting the effect of small molecular compounds of chrysin (C2968) on the expansion of human hematopoietic stem cells by flow cytometry:

[0036] ①Put the collected neonatal umbilical cord blood into a clean and sterile 200mL plasma bottle, add HES according to the volume ratio of hydroxyethyl starch (HES): umbilical cord blood = 4:1, blow and mix well, and let stand at room temperature for 40 Minutes to 1 hour to fully settle the red blood cells in the blood;

[0037] ②Use a 25mL disposable sterile pipet...

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Abstract

The invention relates to application of chrysin in in-vitro expansion of human hematopoietic stem cells. In the application, the concentration of chrysin in an amplification culture medium is 1-10 [mu]M. The invention also provides a human hematopoietic stem cell amplification culture medium and a method for amplifying the human hematopoietic stem cells. The chrysin has a good promotion effect onself-renewal of the human hematopoietic stem cells, and a new method can be provided for clinical treatment in the future.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to the use of chrysin in the in vitro expansion of human hematopoietic stem cells. Background technique [0002] Hematopoietic Stem Cell (HSC) has the ability of self-renewal, multi-directional differentiation, reconstruction of hematopoietic potential and self-repair after injury. Gene therapy and other aspects play an important role. Umbilical cord blood is rich in hematopoietic stem cells, and has the advantages of low immunogenicity, donor safety, wide range of sources, and easy collection. Therefore, umbilical cord blood transplantation has become an excellent alternative source for bone marrow and peripheral blood hematopoietic stem cell transplantation. [0003] However, the proportion of HSCs in umbilical cord blood is only about 0.1% to 1%. Due to many thorny problems such as insufficient quantity, poor quality, and low success rate of engraftment of hematopoietic ste...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0789
CPCC12N5/0647C12N2501/125C12N2501/145C12N2501/999
Inventor 高瀛岱李迎辉杨铭何媚谷佳礼张文姗李亚芳尹晶晶
Owner INST OF HEMATOLOGY & BLOOD DISEASES HOSPITAL CHINESE ACADEMY OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
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