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Method for detecting surface differential membrane protein of mesenchyme stem cells of placenta source

A technology of mesenchymal stem cells and membrane proteins, applied in measuring devices, material analysis through electromagnetic means, instruments, etc., can solve the problems of no clear cell surface markers, etc., and achieve the effect of reducing errors, false positive rates and false negative rates

Active Publication Date: 2013-10-02
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In other words, neither bone marrow, placenta nor umbilical cord MSCS have clear cell surface markers

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0021] The technical solutions of the present invention will be described in detail below in conjunction with specific implementation examples.

[0022] (1) Isolation and culture of mesenchymal stem cells

[0023] Isolation and culture of placental MSCSs:

[0024] Under sterile conditions, the placenta was fully washed with preheated PBS to remove blood stains, cut into pieces as small as possible, digested in 0.1% collagenase type IV, digested at 37°C for 30 minutes, filtered, and washed with PBS Wash, collect the digestion solution and washing solution, centrifuge at 1200 rpm for 10 min, collect the cell pellet, resuspend with 5ml PBS, add to 5ml containing Ficoll-PaqueTM PLUS human lymphocyte separation medium (the relative density is 1.077g / L), carry out Gradient centrifugation (20°C, 2000rpm, 25min). After centrifugation, the buffy coat in the tube was aspirated, washed twice with PBS, and the cells were collected. MSCS-specific medium was resuspended, inoculated ...

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Abstract

The invention relates to a method for detecting differential membrane protein of mesenchyme stem cells, and aims at providing a method for detecting surface differential membrane protein of mesenchyme stem cells of a placenta source. The method comprises the steps of: separating and culturing mesenchyme stem cells, extracting membrane protein from mesenchyme stem cells, obtaining membrane protein gel spectrum by bidirectional fluorescence difference gel electrophoresis, and carrying out mass spectrum analysis and verification. The method utilizes 2D-DIGE (Difference Gel Electrophoresis) for detecting special differential surface markers on the surface of the membrane of the stem cells for the first time, and can compare and analyze protein in different samples of the same gel quantitatively; an interior marker is added in each gel, and DIA and BVA software can automatically calibrate the expression quantity of protein according to the interior marker in each protein point, thus the error between gel of different batches is reduced to the maximum extent, the real change degree of protein is reflected, and the false positive rate and the false negative rate are reduced.

Description

field of invention [0001] The invention relates to a method for detecting differential membrane proteins of stem cells, in particular to a method for detecting differential membrane proteins on the surface of mesenchymal stem cells (Mesenchymal stem cells, MSCSs) derived from placenta and umbilical cord. Background technique [0002] Stem cells (stem cells, SC) are a kind of pluripotent cells with self-replication ability, which can differentiate into various functional cells under certain conditions. Mesenchymal stem cells (MSCSs) are an important member of the stem cell family, originally derived from bone marrow, because of their strong proliferation ability and multi-lineage differentiation potential, hematopoietic support and promotion of stem cell implantation, immune regulation and other characteristics make them useful in tissue engineering, Cell therapy and gene therapy have broad clinical application prospects. In recent years, with the in-depth study of the biolo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/447G01N27/64
Inventor 李兰娟曹红翠俞炯
Owner ZHEJIANG UNIV