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Method for carrying out in-vivo amplification of blood and bone marrow stem cells on newly-born nude mice

A technology of blood stem cells and bone marrow stem cells, applied in the field of expanding blood and bone marrow stem cells in newborn nude mice, to achieve the effect of maintaining basic characteristics, alleviating insufficient cell supply, and reliable cell sources

Active Publication Date: 2014-04-02
深圳市博圣康生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the regulation of its differentiation process, that is, the regulation of the differentiation of stem cells into specific cells, is still a problem that science has not solved well.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The method for expanding blood and bone marrow stem cells in newborn nude mice comprises the following steps:

[0022] (1) Preparation of stem cells

[0023] Use Ficoll density gradient centrifugation and magnetic adsorption separation to separate CD34-positive blood stem cells step by step: take 10ml of fresh human blood, add 50ml of 0.5M EDTA buffer and mix thoroughly, take 38ml of the mixture and add it to a 50ml centrifuge tube Then slowly add 10ml of Ficoll solution from the bottom of the tube, centrifuge at 2000 rpm for 25 minutes at room temperature, carefully absorb about 10ml of the cells in the boundary layer, and then wash the cells once with 10ml of 0.5M EDTA buffer solution. Suspend the magnetic beads of human CD34 antibody in 1ml buffer, incubate on ice for 20 minutes, pass the cell suspension through the filter column in the magnetic field formed by the magnetic separation plate, wash the column twice with 5ml buffer, collect and wash 8 to 10 ml of the l...

Embodiment 2

[0031] The method for expanding blood and bone marrow stem cells in newborn nude mice comprises the following steps:

[0032] (1) Preparation of stem cells

[0033] Use Ficoll density gradient centrifugation and magnetic adsorption separation to separate CD34-positive blood stem cells step by step: take 10ml of fresh human blood, add 50ml of 0.5M EDTA buffer and mix thoroughly, take 38ml of the mixture and add it to a 50ml centrifuge tube Then slowly add 10ml of Ficoll solution from the bottom of the tube, centrifuge at 2000 rpm for 25 minutes at room temperature, carefully absorb about 10ml of the cells in the boundary layer, and then wash the cells once with 10ml of 0.5M EDTA buffer solution. Suspend the magnetic beads of human CD34 antibody in 1ml buffer, incubate on ice for 20 minutes, pass the cell suspension through the filter column in the magnetic field formed by the magnetic separation plate, wash the column twice with 5ml buffer, collect and wash 10ml of the liquid ...

Embodiment 3

[0041] The method for expanding blood and bone marrow stem cells in newborn nude mice comprises the following steps:

[0042] (1) Preparation of stem cells

[0043]Use Ficoll density gradient centrifugation and magnetic adsorption separation to separate CD34-positive blood stem cells step by step: take 10ml of fresh human blood, add 50ml of 0.5M EDTA buffer and mix thoroughly, take 38ml of the mixture and add it to a 50ml centrifuge tube Then slowly add 10ml of Ficoll solution from the bottom of the tube, centrifuge at 2000 rpm for 25 minutes at room temperature, carefully absorb about 10ml of the cells in the boundary layer, and then wash the cells once with 10ml of 0.5M EDTA buffer solution. Suspend the magnetic beads of human CD34 antibody in 1ml buffer, incubate on ice for 20 minutes, pass the cell suspension through the filter column in the magnetic field formed by the magnetic separation plate, wash the column twice with 5ml buffer, collect and wash 10ml of the liquid (...

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Abstract

The invention provides a method for carrying out in-vivo amplification of blood and bone marrow stem cells on newly-born nude mice, which comprises the following steps: preparing separated blood and bone marrow stem cells into a suspension, and performing scalp intravenous injection to carry out in-vivo amplification on newly-born nude mice, wherein the newly-born nude mice survive for 24 hours or below after birth; the variety of the newly-born nude mice is BALB / c-nu; the blood and bone marrow stem cells are subjected to multi-step separation based on a Ficoll density gradient centrifugation method and a magnetic adsorptive separation method; the suspension is prepared by adding normal saline into the blood and bone marrow stem cells; and the concentration of the suspension is 10<6>-10<7> stem cells per ml of normal saline, and the injection dose is 0.05-0.1ml. The invention provides a method for carrying out in-vivo amplification of blood and bone marrow stem cells on animals, which achieves the purposes of keeping the basic characteristics of the stem cells and greatly increasing the quantity of the cells. The invention can provide stem cells having stable performance for clinical cell therapy, can provide a reliable cell source for biomedical research and can effectively relieve the conflict of insufficient cell resources.

Description

technical field [0001] The invention relates to a method for expanding blood and bone marrow stem cells in newborn nude mice. Background technique [0002] Stem cells have the properties of proliferation and differentiation. Under in vitro culture conditions, if no corresponding inhibitory factor (such as leukocyte production inhibitory factor, etc.) is added, the cells will quickly differentiate into other cells of different types and lose the characteristics of stem cells themselves. The regulation of its differentiation process, that is, the regulation of the differentiation of stem cells into specific cells, is still a problem that science has not yet solved well. How to keep stem cells in an undifferentiated state, that is, still have the basic characteristics of stem cells and allow the cells to proliferate in large quantities, has become a major problem in this research field. Contents of the invention [0003] The technical problem to be solved by the present inv...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0789
Inventor 杨寅柯高红杨登科
Owner 深圳市博圣康生物科技有限公司
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