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A kind of in vitro expansion medium of umbilical cord blood hematopoietic stem cells and its application

A technology for in vitro expansion of hematopoietic stem cells, applied in the field of in vitro expansion medium for umbilical cord blood hematopoietic stem cells, can solve the problems of low cell proliferation rate, low implantation ability and hematopoietic reconstruction ability, and achieve high proliferation rate, implantation Improve the ability and rebuilding ability of hematopoietic system, promote the effect of application

Active Publication Date: 2018-07-10
广东美赛尔细胞生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to solve the defects of low cell proliferation rate in the in vitro expansion medium of umbilical cord blood hematopoietic stem cells in the prior art, and low engraftment ability and hematopoietic reconstruction ability of the umbilical cord blood hematopoietic stem cells cultured in vitro, this The object of the invention is to provide a umbilical cord blood hematopoietic system with a high proliferation rate of hematopoietic stem cells, which can significantly improve the implantation ability of the hematopoietic stem cells transplanted into the recipient and the reconstruction ability of the hematopoietic system, and can better maintain the stemness of the stem hematopoietic cells. Stem cell in vitro expansion medium to solve the above problems

Method used

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  • A kind of in vitro expansion medium of umbilical cord blood hematopoietic stem cells and its application
  • A kind of in vitro expansion medium of umbilical cord blood hematopoietic stem cells and its application
  • A kind of in vitro expansion medium of umbilical cord blood hematopoietic stem cells and its application

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Effect test

Embodiment 1

[0026] Example 1. Effects of Medium Containing Different Concentrations of Berberine on the Proliferation of CD34+CD38-Lin- Cells in Vitro

[0027] 1. Test materials: Umbilical cord blood was collected from the Department of Obstetrics and Gynecology of Huaqiao Hospital, and the mononuclear cell isolation kit was purchased from Beijing Suolaibao Technology Co., Ltd. (article number: P9260).

[0028] 2. Test method: the collected umbilical cord blood was separated from mononuclear cells with a mononuclear cell isolation kit to obtain mononuclear cells from umbilical cord blood, and then CD34+CD38-Lin- cells were separated by immunomagnetic bead cell sorting, and used Cell purity was identified by flow cytometry.

[0029] Release berberine into 2, 5, 7, and 10 ug / ml drug solutions with IMEM basal medium, and separate the cells of P1 generation at 10 4Cells / mL were inoculated in 96-well plates, and the in vitro expansion medium of umbilical cord blood hematopoietic stem cells co...

Embodiment 2

[0031] Embodiment 2. In vitro expansion medium of umbilical cord blood hematopoietic stem cells

[0032] The medium for in vitro expansion of umbilical cord blood hematopoietic stem cells, including IMEM basal medium, also includes the following components and their concentrations: fetal bovine serum 0.05mg / mL, thrombopoietin 15ng / mL, stem cell factor 100ng / mL, human FMS-like Tyrosine kinase 3 ligand 33ng / ml, interleukin-1 0.2ng / ml, interleukin-6 0.2ng / ml, stem cell growth factor 0.2ng / ml and berberine 2ug / ml.

[0033] Preparation method: Add fetal bovine serum, thrombopoietin, stem cell factor, human FMS-like tyrosine kinase 3 ligand, interleukin-1, interleukin-6, stem cell growth factor and small Berine, mixed evenly, passed through membrane to sterilize.

Embodiment 3

[0034] Embodiment 3, in vitro expansion medium of umbilical cord blood hematopoietic stem cells

[0035] The medium for in vitro expansion of umbilical cord blood hematopoietic stem cells, including IMEM basal medium, also includes the following components and their concentrations: fetal bovine serum 0.1mg / mL, thrombopoietin 25ng / mL, stem cell factor 120ng / mL, human FMS-like Tyrosine kinase 3 ligand 45ng / ml, interleukin-1 0.3ng / ml, interleukin-6 0.6ng / ml, stem cell growth factor 0.5ng / ml and berberine 5ug / ml.

[0036] The preparation method is similar to Example 2.

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Abstract

The invention belongs to the technical field of stem cells, and relates to an in vitro expansion medium of umbilical cord blood hematopoietic stem cells and an application thereof. The in vitro expansion medium includes IMEM basal medium, and also includes the following components: serum, thrombopoietin, stem cell factor, FMS-like tyrosine kinase 3 ligand, interleukin-1, interleukin-6, stem cell growth factor and Berberine. The in vitro expansion medium of umbilical cord blood hematopoietic stem cells provided by the present invention has the advantage of high proliferation rate of hematopoietic stem cells, and can significantly improve the implantation ability of hematopoietic stem cells transplanted into the recipient and the reconstruction ability of the hematopoietic system, and can better maintain The characteristic of hematopoietic stem cells is an ideal in vitro expansion medium for umbilical cord blood hematopoietic stem cells.

Description

technical field [0001] The invention belongs to the technical field of stem cells, and relates to an in vitro expansion medium of umbilical cord blood hematopoietic stem cells and an application thereof. Background technique [0002] Hematopoietic stem cells (HSCs) are adult stem cells in the blood system. They are a heterogeneous population with the ability of long-term self-renewal and the potential to differentiate into various mature blood cells. Because hematopoietic stem cells have the potential of multi-directional differentiation, transplanting hematopoietic stem cells into the body is an effective method for the treatment of leukemia, lymphoma and other malignant tumors, metabolic diseases, autoimmune diseases and congenital immune deficiencies that seriously endanger human health. alleviating the suffering of patients. However, due to the extremely small number of hematopoietic stem cells in the body, the clinical application of hematopoietic stem cells is greatly...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0789
Inventor 王一飞程江王巧利廖晓凤刘秋英
Owner 广东美赛尔细胞生物科技有限公司
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