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In-vitro amplification method for cord blood hematopoietic stem cells

A technology for in vitro expansion of hematopoietic stem cells, which is applied in the field of in vitro expansion of umbilical cord blood hematopoietic stem cells, can solve the problems of limited quantity and restriction of adult application factors, and achieve low cost and good use effect

Inactive Publication Date: 2015-11-04
GUIZHOU BEIKE FACTORR BIOTECH CO LTD
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  • Summary
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  • Description
  • Claims
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Problems solved by technology

However, according to the minimum standards required for bone marrow and peripheral blood transplantation (1-2×10 8 cell / kg body weight or 0.5-1.0×10 6 CD34 + cells / body weight), the amount of stem cells contained in every 100ml of cord blood can only meet the needs of children not greater than 30kg, while the amount of stem cells for adults is limited and limited

Method used

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Embodiment Construction

[0020] Embodiments of the present invention: a method for in vitro expansion of umbilical cord blood hematopoietic stem cells, comprising the following steps:

[0021] 1) Mix cord blood and serum-free medium at a volume ratio of 1:1; separate mononuclear cells in cord blood with lymphocyte separation medium with a specific gravity of 1.077, and separate lymphocyte separation medium and cord blood with a 50ml centrifuge tube The mixture was mixed with a volume ratio of 10:35, and centrifuged at 2000 rpm for 20 minutes at room temperature;

[0022] 2) Aspirate the mononuclear cell suspension, wash the mononuclear cell suspension with 30ml saline twice, centrifuge at 1000rpm for 10 minutes each time, and discard the supernatant;

[0023] 3) 1 / 3 of the obtained mononuclear cells were cultured in vitro, and the other 2 / 3 of the cells were stored in liquid nitrogen;

[0024] 4) Serum-free medium for in vitro cell culture, add stem cell growth factor 100ng / ml, thrombopoietin TPO 100...

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Abstract

The invention provides an in-vitro amplification method for cord blood hematopoietic stem cells. According to the invention, culture process is optimized and screened, so cultured cord blood hematopoietic stem cells can meet demands of populations needing a great number of cord blood hematopoietic stem cells, have longer life compared with hematopoietic stem cells obtained after bone marrow transplantation and have low incidence rate of graft-versus-host disease. The culturing method provided by the invention is simple and easily practicable and has low cost and good application effect.

Description

technical field [0001] The invention relates to the technical field of biological sciences, in particular to an in vitro expansion method of umbilical cord blood hematopoietic stem cells. Background technique [0002] In recent years, people have tried to use human umbilical cord blood hematopoietic stem cells (HSC) instead of adult bone marrow for transplantation to treat some malignant diseases. However, according to the minimum standards required for bone marrow and peripheral blood transplantation (1-2×10 8 cell / kg body weight or 0.5-1.0×10 6 CD34 + Cells / body weight), the amount of stem cells contained in every 100ml of cord blood can only meet the needs of children not greater than 30kg, while the application to adults is limited due to the limited amount. Contents of the invention [0003] The purpose of the present invention is to provide a method for in vitro expansion of umbilical cord blood hematopoietic stem cells. The HSCA expanded by this method can not on...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0789
Inventor 边曙光
Owner GUIZHOU BEIKE FACTORR BIOTECH CO LTD
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