Culture method and culture medium for in vitro amplification of hematopoietic stem cells

A technology for hematopoietic stem cells and a culturing method is applied in the field of culture methods and culture medium for expanding hematopoietic stem cells in vitro, and can solve the problems of limited sources of hematopoietic stem cells, hindering wide application, and low quantity.

Inactive Publication Date: 2019-04-26
南京温博生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the source of hematopoietic stem cells is very limited and the number is extremely low, which greatly hinders their widespread application
In vitro culture and expansion is an important means to increase the number of hematopoietic stem cells, however, conventional in vitro culture and expansion methods are not satisfactory for the expansion of hematopoietic stem cells

Method used

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  • Culture method and culture medium for in vitro amplification of hematopoietic stem cells
  • Culture method and culture medium for in vitro amplification of hematopoietic stem cells
  • Culture method and culture medium for in vitro amplification of hematopoietic stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: In vitro expansion method of hematopoietic stem cells

[0025] 1. Experimental materials

[0026] Mice were healthy male C57BL / 6 mice aged 6-8 weeks, weighing 22±2g; PBS buffer was purchased from Shanghai Shushibiology; Ficoll lymphocyte separation liquid was purchased from Tianjin Haoyang Biological Products; Anti-Sca-1 -PE, Anti-PEMicroBeads, and MS separation columns were purchased from Miltenyi Company in Germany; IMDM culture medium and fetal bovine serum were purchased from Hyclone Company in the United States; BCA kit was purchased from Beyontian; rabbit anti-mouse p18 monoclonal antibody was purchased from Santa Cruz in the United States Biotechnology company, HRP-labeled secondary antibody was purchased from Solarbio. The purity of wogonin, dehydrated icariin, honokiol and anisrolactone is not less than 95%.

[0027] 2. Experimental method

[0028] 1. Preparation of mouse bone marrow mononuclear cells (BMNCs)

[0029] Mice were sacrificed under a...

Embodiment 2

[0070] Embodiment 2: In vitro expansion medium of hematopoietic stem cells

[0071] According to Example 1, adding a p18 protein inhibitor during hematopoietic stem cell culture can increase the expansion speed of hematopoietic stem cells, and an appropriate amount of p18 protein inhibitor can be added to the IMDM medium, such as wogonin, dehydrated epimeditin, and thick Parkol or anisolactone, made of commercial medium for the in vitro expansion of hematopoietic stem cells.

[0072] The above examples show that adding a p18 protein inhibitor during hematopoietic stem cell culture can increase the proliferation rate of hematopoietic stem cells. The p18 protein inhibitor can be an RNA fragment, peptide or small molecule compound that inhibits the expression of p18 protein in hematopoietic stem cells. The RNA fragment is The interference RNA, the peptide is a polypeptide or an oligopeptide, and the small molecular compound is wogonin, dehydroicariin, honokiol or aniselide. The ...

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Abstract

The invention discloses a culture method and a culture medium for in vitro amplification of hematopoietic stem cells. By using the method, the result that the hematopoietic stem cell amplification speed can be accelerated through the addition of a proper number of p18 protein inhibitors during hematopoietic stem cell culture is discovered. The p18 protein inhibitors can be p18 protein expression inhibition RNA fragments, peptide or micromolecular compounds in the hematopoietic stem cells; the RNA fragments are interfering RNA; the peptide is polypeptide or oligopeptides; the micromolecular compounds are wogonin, anhydroicaritin and magnolol or pimpinellin. The p18 protein inhibitors can be used for preparing the hematopoietic stem cell vitro amplification culture medium.

Description

technical field [0001] The invention belongs to the biological field and relates to the cultivation of hematopoietic stem cells, in particular to a culture method and culture medium for expanding hematopoietic stem cells in vitro. Background technique [0002] Hematopoietic stem cells (HSCs) exist in very low numbers in mammalian bone marrow, fetal liver, peripheral blood and umbilical cord blood. They are stem cells with high self-renewal ability and multi-directional differentiation potential, and are the precursor cells of all mature blood cells. . Hematopoietic stem cell transplantation is an important method for the treatment of malignant blood diseases, genetic diseases and immunodeficiency diseases, and has played an important role in the treatment of many fatal diseases. However, the sources of hematopoietic stem cells are very limited and the quantity is extremely low, which greatly hinders their widespread application. In vitro culture expansion is an important m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0789
CPCC12N5/0647C12N2501/405
Inventor 不公告发明人
Owner 南京温博生物科技有限公司
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