High-efficiency method for inducing human mesenchymal stem cells to differentiate into cartilage

A technique of mesenchymal stem cells and induction of differentiation, applied in the field of stem cells, can solve the problems of non-pellet formation, reduction of stem cells, irregular shape of cell pellets, etc., and achieve the effect of preventing wrinkles and being in good condition

Pending Publication Date: 2021-04-16
朱灏
View PDF3 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this application adds fetal bovine serum albumin to the cell culture medium cultured in vitro, and the biological activity of fetal bovine serum is relatively high. Higher activity, it is easy to promote stem cells in the process of induction and differentiation, and the shape of cell pellets after cell differentiation is irregular or not into pellets; and the faster induction of differentiation into cartilage is likely to reduce the quality of stem cells differentiated into cartilage In addition, during the use of fetal bovine serum, due to its complex internal components and being easily affected by some external factors, it is easy to adversely affect the quality of the medium, such as containing viruses, which seriously affects the proliferation and cultivation of stem cells. The effect of chondrogenic differentiation

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
  • High-efficiency method for inducing human mesenchymal stem cells to differentiate into cartilage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] This embodiment provides a high-efficiency method for inducing differentiation of human mesenchymal stem cells into cartilage. The steps include the following steps: (1) 1 mL of serum substitute EliteGro TM Prepare the cell culture medium with 19mL of mesenchymal stem cell serum-free medium NutriStemXF Medium, and store it at 2°C for use after preparation; Mix was prepared as a chondrogenic differentiation medium, and after the preparation was completed, it was stored at 2°C for later use; (3) Mesenchymal stem cells were resuspended in cell culture medium, and the density of mesenchymal stem cells was adjusted to 1x10 5 / 10μl, and use the 96-well plate to inoculate the mesenchymal stem cells, the inoculation density is 1x10 5 / 10μl / well; (4) After inoculation, place at 37°C, 5% CO 2 After standing in the incubator with high concentration for 2 hours, add 0.1 mL of cell culture medium; (5) after standing for 24 hours, replace the medium with chondrogenic differentiation...

Embodiment 2

[0059] The specific implementation of this example is the same as that of Example 1, except that: the serum-free medium NutriStem XF Medium for mesenchymal stem cells is 20 mL.

Embodiment 3

[0061] The specific implementation method of this example is the same as that of Example 1, except that the Chondrogenic Differentiation Basal Medium of the basal medium is 16 mL.

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 discloses a method for inducing human mesenchymal stem cells to differentiate into cartilage in vitro. The method comprises the following steps: (1) preparing a cell culture medium, and after preparation, preserving at 2-8 DEG C for later use; (2) preparing a compound cartilage induced differentiation culture medium, and after preparation, storing at 2-8 DEG C for later use; (3) resuspending the mesenchymal stem cells by using a cell culture medium, adjusting the density of the mesenchymal stem cells, and inoculating the mesenchymal stem cells by using a 96-well plate; (4) after the inoculation is finished, placing the culture medium in a CO2 incubator, standing for 2-3 hours, and supplementing a cell culture medium for subsequent culture; and (5) after standing for 24 hours, replacing the culture medium with a cartilage induced differentiation culture medium, and carrying out cartilage induced differentiation. The method has good stem cell culture and protection effects, stem cells obtained after cartilage differentiation are all pellet-shaped and regular in shape, obvious stem cell damage is avoided, the cartilage time period efficiency is high, and the method is suitable for being popularized in the field of stem cells and has wide development prospects.

Description

technical field [0001] The invention relates to the field of stem cells, in particular to a method for inducing differentiation of human mesenchymal stem cells into cartilage with high efficiency. Background technique [0002] The differentiation of stem cells is the selective activation or differential expression of some genes, thereby controlling the cell phenotype and the specific distribution of proteins. The differentiation of mesenchymal stem cells into specific cell types mainly depends on the activation of genes, and the types and concentrations of various factors in the extracellular microenvironment are important factors for gene activation. At present, the methods of inducing differentiation of mesenchymal stem cells into chondrocytes in vitro mainly include: (1) traditional chemical inducer method; (2) growth factor induction method; (3) combination of growth factor and chemical inducer; (4) co-culture Or use the conditioned medium close to the physiological sta...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077C12N5/0775
Inventor 朱灏
Owner 朱灏
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap