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

Preparation method and application of parietal decidua subtotipotent stem cells

A technology of subtotipotent stem cells and decidua parietal, applied in the field of preparation of subtotipotent stem cells in parietal decidua, can solve the problems of limiting the application of subtotipotent stem cells, hindering the extraction of subtotipotent stem cells, and low activity of subtotipotent stem cells, and achieving strong differentiation ability , the effect of rich sources and large quantities

Active Publication Date: 2022-04-12
GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are some obstacles in the extraction of sub-totipotent stem cells. On the one hand, most of the existing sub-totipotent stem cells are extracted from human placenta, which has certain ethical problems. On the other hand, the sub-totipotent stem cells obtained by the existing extraction methods have low activity. , Differentiation ability is limited, which limits the application of subtotipotent stem cells in clinical treatment

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
  • Preparation method and application of parietal decidua subtotipotent stem cells
  • Preparation method and application of parietal decidua subtotipotent stem cells
  • Preparation method and application of parietal decidua subtotipotent stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] A method for preparing subtotipotent stem cells in decidua wall, comprising the following steps:

[0033] (1) Pretreatment: Take out the parietal decidua tissue from the collection bag, wash it with tissue cleaning solution until there is no blood residue, then put the cleaned placenta tissue into a 50mL sterile centrifuge tube, centrifuge at 1200rpm for 5min, removing the supernatant; wherein, the cleaning solution includes: physiological saline, red blood cell lysate with a volume fraction of 50%, gentamicin sulfate at 10 μg / ml, and amphotericin B at 10 μg / ml.

[0034] (2) Digestion: Cut the cleaned parietal decidua tissue into 0.1-1cm with sterile direct scissors 3Add an equal volume of tissue digestion solution preheated to 37°C, mix well, shake and digest in a constant temperature shaker at 37°C for 2 hours at a speed of 200r / min, centrifuge quickly and transiently after digestion, collect the supernatant, and pour Add normal saline to the precipitate to resuspend...

experiment example 1

[0046] The P0 generation cell and the P3 generation cell count of embodiment and comparative example gained, the result is as shown in table 1:

[0047] Table 1 Subtotipotent stem cell count table of parietal decidua

[0048]

experiment example 2

[0050] Identification of osteoblast differentiation ability:

[0051] Subtotipotent stem cells of the P3 generation in Example 1 were divided into 2×10 4 Inoculate in a 24-well plate at a concentration of / mL, 1 mL per well, and replace the osteoblast differentiation induction medium every 3 days. Semethasone 10nM / mL, β-glycerol phosphate 10M / mL, antibiotic 1wt%, cytokine IL-β 25ng / mL, ascorbic acid 105mM / mL, isoflavone 10μg / mL. Cultivate until the 14th day, discard the medium, wash twice with PBS, fix with 4% paraformaldehyde for 20 minutes, and stain with Alizarin Red. After staining for 20 minutes, wash twice with normal saline to observe the formation of calcium nodules Obviously, it shows that the subtotipotent stem cells in Example 1 can effectively differentiate into osteoblasts.

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 provides a preparation method and application of decidua parietal subtotipotent stem cells, and relates to the technical field of cell engineering. The preparation method of the present invention comprises the following steps: pretreatment: cleaning the parietal decidua group, centrifuging, and removing the supernatant; digestion: cutting the parietal decidua tissue into pieces, adding tissue digestion liquid for digestion, and separating the supernatant and precipitation after digestion , resuspending the precipitate to obtain a suspension; the tissue digestion solution includes Tryple enzyme, normal saline, type I collagenase and type II collagenase, the volume concentration of the Tryple enzyme is 40% to 60%, and the volume concentration of type I collagenase The mass concentration is 0.8-1.2mg / mL, and the mass concentration of type II collagenase is 0.8-1.2mg / mL; Separation: add lymphocyte separation liquid to the suspension, centrifuge, separate the mononuclear cells after layering, centrifuge, Keep the precipitate, add culture medium, and mix well to obtain parietal decidua subtotipotent stem cell separation medium. The invention can successfully extract subtotipotent stem cells from parietal decidua tissue, and has good cell activity and strong differentiation ability.

Description

technical field [0001] The invention relates to the technical field of cell engineering, in particular to a preparation method and application of parietal decidua subtotipotent stem cells. Background technique [0002] Stem cells are a type of cells with self-replication, renewal and multi-directional differentiation potential, which can be differentiated into various tissues and organs. Stem cells have become a research hotspot in the field of life sciences. Scientists have discovered that there is a type of primitive stem cell population in various adult tissues. They are different from pluripotent stem cells such as hematopoietic stem cells and neural stem cells. Pluripotent stem cells can only differentiate into cells of a specific germ layer, while these primitive stem cells can differentiate into different germ layers. tissue cells. This kind of primitive stem cells is also different from embryonic stem cells. During the growth and development of individuals, they gra...

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): C12N5/073C12N5/077C12N5/071A01N1/02
CPCC12N5/0605C12N5/0654C12N5/0682C12N5/0625A01N1/0221C12N2509/00C12N2509/10C12N2506/025C12N2501/39C12N2500/42C12N2500/38C12N2501/392C12N2501/2302C12N2501/15C12N2501/11C12N2501/135
Inventor 刘小翠李静静赵蓝孙灿兴江嘉豪褚一凡
Owner GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
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