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Method for promoting activation of umbilical cord stem cells and synergistically intervening knee osteoarthritis through platelet lysate

A technology for knee osteoarthritis and umbilical cord stem cells, applied in anti-inflammatory agents, bone diseases, drug combinations, etc. Problems such as the lack of verification of the auxiliary effect, to achieve the effect of improving viability and biological function

Pending Publication Date: 2020-10-13
ZHEJIANG CHINESE MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the prior art, there have been reports on the expansion and culture of umbilical cord stem cells by platelet lysates. A method for culturing umbilical cord mesenchymal stem cells without animal serum and allogeneic proteins has been disclosed before, but this technology does not provide platelet lysates. The specific cell molecular mechanisms related to the proliferation, chemotaxis and regeneration of umbilical cord stem cells. In addition, in the existing research reports, the auxiliary effect of platelet lysates on stem cell transplantation treatment is still lacking in animal experiments. To solve the above problems, the present invention adopts CCK -8 method, flow cytometry identification, ELISA method, scratch test, Transwell and RT-PCR method, optimize the combination of platelet lysate and umbilical cord stem cells, and explore the mechanism of growth factor-based platelet lysate acting on umbilical cord stem cells

Method used

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  • Method for promoting activation of umbilical cord stem cells and synergistically intervening knee osteoarthritis through platelet lysate
  • Method for promoting activation of umbilical cord stem cells and synergistically intervening knee osteoarthritis through platelet lysate
  • Method for promoting activation of umbilical cord stem cells and synergistically intervening knee osteoarthritis through platelet lysate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Preparation of platelet lysate

[0024] Collect peripheral blood with a blood collection tube containing 3.2% anticoagulant (sodium citrate), let it stand at room temperature for more than 30 minutes, centrifuge at 210 g for 10 minutes, take the upper and middle layers, discard the lower layer of plasma, and centrifuge at 210 g for 5 minutes , to remove the remaining red blood cells. Then put it into liquid nitrogen or -80°C refrigerator for repeated freeze-thaw lysis (3 cycles), or ultrasonic lysis to obtain platelet lysate (PL).

Embodiment 2

[0025] Embodiment 2: the mensuration of platelet lysate purity

[0026] The PL extracted in Example 1 was identified by flow cytometry, with the addition of 0.9% normal saline and 10% PL. The result is as figure 1 As shown in A, the positive expression rate of CD41a in PL is 99.11%, indicating that the prepared human platelet lysate PL has a high purity.

Embodiment 3

[0027] Example 3: Determination of cytokine content in platelet lysate

[0028] The content of cytokines in PL was detected by ELISA method, and the additives were 0.9% normal saline and 10% PL. The result is as figure 1 As shown in B, the content of PDGF is 26.25±0.48 ng / ml, the content of TGF-β is 7.62±1.29 ng / ml, the content of IFG-1 is 5.05±0.15 ng / ml, the content of EGF is 1.39±0.08 ng / ml, the content of FGF It was 0.33±0.01 ng / ml.

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Abstract

The invention discloses a method for promoting activation of umbilical cord stem cells and synergistically intervening knee osteoarthritis through a platelet lysate, and belongs to the technical fieldof umbilical cord stem cell application. The invention provides an application of the platelet lysate in preparation of an umbilical cord stem cell adjuvant and an application of the platelet lysate,in cooperation with the umbilical cord stem cells, in preparation of knee osteoarthritis treatment drugs. Through preparation of the platelet lysate, the purity of the platelet lysate, the content ofcytokines, the proliferation activity of the umbilical cord stem cells, the cycle progress, the cell cycle related gene expression, the scratch repair capacity, the chemotactic migration capacity andthe chemotactic migration related gene expression are measured, and rat behavioristics and histopathology observation tests are carried out. It is verified that the platelet lysate is suitable for being used as the umbilical cord stem cell adjuvant, and the mechanism that the platelet lysate based on growth factors acts on the umbilical cord stem cells and the synergistic application of the platelet lysate in knee osteoarthritis treatment are further defined.

Description

technical field [0001] The invention belongs to the technical field of application of umbilical cord stem cells, and in particular relates to a method for promoting the activation of umbilical cord stem cells by platelet lysates and synergistically intervening in knee osteoarthritis. Background technique [0002] Mesenchymal stem cells (MSCs) have self-renewal ability, multi-lineage differentiation potential, paracrine effect and immune regulation function, which can promote the regeneration of damaged tissue, and are an important therapeutic tool for cell therapy and regenerative medicine. Mesenchymal stem cells come from a wide range of sources, such as bone marrow, adipose tissue, and menstrual blood. The umbilical cord has long been considered a medical waste, but compared with stem cells derived from bone marrow, umbilical cord stem cells (human umbilical cord-derived MSCs, huc-MSCs) have higher plasticity and fewer ethical constraints, so the umbilical cord is a A ver...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/51A61K35/19A61P19/02A61P29/00A61P19/08
CPCA61K35/51A61K35/19A61P19/02A61P29/00A61P19/08A61K2300/00
Inventor 单乐天
Owner ZHEJIANG CHINESE MEDICAL UNIVERSITY