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Method of separating blood vessel endothelium precursor cell from marrow and cultivating in vitro

A precursor cell, in vitro culture technology, applied in artificial cell constructs, tissue culture, animal cells, etc., can solve problems such as limited number

Inactive Publication Date: 2007-02-28
SHANGHAI HIGH SCHOOL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, vascular endothelial precursor cells exist in extremely limited numbers in peripheral blood

Method used

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Embodiment Construction

[0021] The present invention will be described in detail below with the embodiment of isolating vascular endothelial precursor cells from the bone marrow of Wistar rats and culturing them in vitro:

[0022] A method for isolating vascular endothelial precursor cells from bone marrow and culturing them in vitro, comprising the following steps:

[0023] 1. Harvesting bone marrow cells: Wistar rats were killed by cervical dislocation, soaked in 75% ethanol for about 15 minutes for disinfection. Ultra-clean table, disinfection of surgical instruments. The rat hindlimb was separated from the hip joint, the femur and tibia were exposed, and the blood-stained villi were removed. The scissors cut off the epiphysis at both ends, exposing the bone marrow cavity. Use a 10ml syringe to draw the EBM-2 culture solution containing 100IU / ml heparin, inject it into the bone marrow cavity through a needle, collect the washed bone marrow, blow the harvested bone marrow repeatedly with a syring...

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Abstract

The invention discloses the method for separating vessel esoderma fore-body cell from bone marrow and extracorporal culturing. The method comprises the following steps: 1 getting marrow cell; 2 collecting individual cell; 3 making EBM-2 cell culture liquid; 4 cell culturing; 5 passage culturing. The bFGF and VEGF are added in EBM-2 cell culture liquid to improve proliferation of vessel esoderma fore-body cell. So the invention can get many vessel esoderma fore-body cells in a short time.

Description

technical field [0001] The invention relates to a method for separating and culturing cells, especially a method for isolating vascular endothelial precursor cells from bone marrow and culturing them in vitro. Background technique [0002] Stem / progenitor cells derived from autologous bone marrow can proliferate in large quantities, are not easy to age, and can be induced in vitro to differentiate into multiple directions. They have become research hotspots in the fields of cell transplantation and tissue engineering in recent years. The maintenance of biological activity of bone marrow stem / progenitor cells in vivo is regulated by a series of complex factors, for example, EPCs present in bone marrow are closely related to hematopoietic stem cells and bone marrow stromal cells. Hematopoietic stem cells, differentiated osteoblasts and osteoclasts in the bone marrow secrete various pro-angiogenic factors, such as VEGF, to promote the proliferation, recruitment and mobilization...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/08C12N5/071
Inventor 林昕
Owner SHANGHAI HIGH SCHOOL
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