Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Identification and application of human amniotic mesenchymal stem cell exocrine protein POSTN

An amniotic membrane mesenchymal and stromal cell technology, which can be used in medical preparations containing active ingredients, applications, peptide/protein components, etc., and can solve the problems that POSTN application research has not been in-depth and POSTN has been identified.

Active Publication Date: 2021-08-17
中国医科大学
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, POSTN has not been identified in the exocrine protein of human amniotic mesenchymal stem cells, and there is no related identification method report. At the same time, the application research on POSTN has not been in-depth. There is no report on the relationship between the effects of the drug and the clinical treatment of male infertility and the preservation of male fertility

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Identification and application of human amniotic mesenchymal stem cell exocrine protein POSTN
  • Identification and application of human amniotic mesenchymal stem cell exocrine protein POSTN
  • Identification and application of human amniotic mesenchymal stem cell exocrine protein POSTN

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] 1. Harvest of human amniotic mesenchymal stem cell conditioned medium

[0024] Materials and Methods:

[0025] Protease inhibitor (Biyuntian)

[0026] trypsin BI

[0027] Ultrafiltration centrifuge tubes (Merck Millipore; Amicon Ultra-50, Ultracel-3k)

[0028] BCA kit (Biyuntian)

[0029] Type IV collagenase (sigma)

[0030] Osteogenic / Adipogenic / Chondrogenesis Kit (Saiye Bio)

[0031] (1) The amniotic membrane mesenchymal stem cells are obtained by the following methods:

[0032] The amniotic tissue of healthy caesarean section was taken, soaked in normal saline for 30 min, and washed repeatedly with PBS to remove mucus. Use ophthalmic surgical scissors to cut the amniotic membrane into small pieces of about 1mm × 1mm, add 10ml of 0.25% trypsin-EDTA, digest at 37°C for 30 minutes, add serum-containing culture medium, centrifuge to remove the supernatant, and repeat the digestion twice, and then use Wash 2-3 times with PBS to remove as much epithelial cells as po...

Embodiment 2

[0040] Mass spectrometry analysis of exocrine proteins from amniotic mesenchymal stem cells in conditioned medium. Protein annotation was analyzed by GeneOntology (GO). GO analysis is an important bioinformatics analysis method used to describe various characteristics of genes and gene expression products. GO annotations are divided into three categories: biological process, cellular composition and molecular function. These three classifications explain the biological functions of proteins from different perspectives. We performed statistics on the distribution of hAMSC-secreted proteins in GO secondary annotations. The results showed that exocrine proteins in hAMSCs-CM were involved in biological processes such as cell proliferation, differentiation, metabolism, immunity, and signaling pathways. The highly expressed protein POSTN was screened out. ELISA verifies the highly expressed screening proteins in amniotic mesenchymal stem cells.

[0041] Materials and Methods:

...

Embodiment 3

[0046] MTS and EDU experiments found that the protein can effectively promote the proliferation of spermatogonial stem cells.

[0047] The proliferation of cells was detected by MTS assay, according to 2.5 × 10 per well. 4 cells were seeded in 96-well plates. After 24h, different concentrations of POSTN were added to the corresponding wells. Every 24h, add 10µl MTS to the wells to be tested, then incubate at 37°C for 2h, and use a microplate reader to measure the value at a wavelength of 490nm.

[0048] In order to determine whether the two candidate proteins POSTN in hAMSCs-CM are related to cell proliferation, different concentrations (0, 10, 50, 100, 200 ng / mL) of pure POSTN protein were added to the cell proliferation medium, respectively. Cell proliferation was detected by MTS method at 0, 24, 48, 72, and 96 h. like Figure 4 As shown, the results showed that the higher the concentration of POSTN, the stronger the promoting effect on cell proliferation. From 48 hours...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to application of human amniotic mesenchymal stem cell exocrine protein POSTN in preparation of a product for promoting spermatogonial stem cell proliferation and an identification method of the exocrine protein POSTN. The identification method comprises the following steps: collecting a human amniotic mesenchymal stem cell conditioned medium; and carrying out mass spectrometry on the exocrine protein of the amniotic mesenchymal stem cells in the conditioned culture medium, and screening out the high-expression protein POSTN. The exocrine proteome of the amniotic mesenchymal stem cells is identified, the results are analyzed and integrated, the spermatogonial stem cell proliferation positive factor POSTN is screened out, and a detailed foundation and a new thought are provided for subsequent research on amniotic stem cell paracrine effects. The POSTN provides a new cell factor for in-vitro proliferation of spermatogonial stem cells, and provides a direction for better excavating the characteristics of exocrine proteins of amniotic stem cells.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the identification and application of human amniotic mesenchymal stem cell exocrine protein-periostin (POSTN). Background technique [0002] Mesenchymal stem cells (MSCs) have been shown to secrete various autocrine / paracrine factors, including growth factors and cytokines, which play a major role in cell therapy. Human amniotic membranes are often discarded as biological waste after childbirth and become an important source of amniotic mesenchymal stem cells (hAMSCs). In addition, it has many other advantages such as anti-inflammatory, antibacterial, anti-angiogenic properties and low immunogenicity. HAMSC-secreted factors for cell-free therapy of acute brain injury. We found many proteins related to cell proliferation in hAMSCs conditioned medium (hAMSCs-CM). [0003] Periostin (POSTN) is an extracellular matrix protein composed of 836 amino acids and about 90kD in size. POSTN ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): A61K38/17A23L33/17A61P15/00G01N15/14G01N27/62G01N33/68
CPCG01N33/6848A61K38/1709A61P15/00A23L33/17G01N27/62G01N15/14A23V2002/00A23V2200/30A23V2250/542
Inventor 庞希宁李彩虹
Owner 中国医科大学
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com