Human umbilical cord blood mesenchymal stem cell extract and preparation method and application thereof

A technology of stem cells and human umbilical cord blood, which is applied in the field of biomedical stem cells and tissue engineering, can solve the problem of single active ingredient and achieve the effect of accelerating the reproduction rate and increasing the yield

Inactive Publication Date: 2019-01-04
湖南艾佳生物科技股份有限公司
View PDF8 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the above-mentioned technical problem that the active ingredient obtained from human umbilical cord blood mesenchymal stem cel

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
  • Human umbilical cord blood mesenchymal stem cell extract and preparation method and application thereof
  • Human umbilical cord blood mesenchymal stem cell extract and preparation method and application thereof
  • Human umbilical cord blood mesenchymal stem cell extract and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0035] Example 1 Isolation and culture of human umbilical cord blood mesenchymal stem cells

[0036] 1.1 Cord blood collection

[0037] With the assistance of the obstetrics and gynecology hospital that has a long-term cooperation with our company, cord blood will be collected from cesarean women who are healthy under 30 years old, have no genetic diseases, no infectious diseases, and have negative anti-HIV and hepatitis B serum markers. . After collection, the cord blood was placed in an ice box and immediately transported to the laboratory for separation and processing. Among them, the parturients involved knew and agreed to the experimental protocol of the separated cord blood.

[0038] 1.2 Isolation and culture of mesenchymal stem cells from cord blood

[0039] The collected umbilical cord blood is diluted in an equal volume with phosphate buffered saline (PBS for short), and then the lymphatic separation solution is used for density gradient centrifugation to collect the albugi...

Example Embodiment

[0041] Example 2 Identification of cell types of isolated human umbilical cord blood mesenchymal stem cells

[0042] 2.1 Take the umbilical cord blood mesenchymal stem cells cultured in vitro to the third generation, digest, centrifuge, and adjust the cell density to 1x106 cells / ml with PBS, and divide them into multiple tubes.

[0043] 2.2 Absorb the following antibodies: ITC-anti-human-IgG FC (purchased from Biolegend, item number 409309), FITC-anti-hunman-CD44 (purchased from Biolegend, item number 338803), FITC-anti-hunman-CD105 (purchased from Biolegend , Article number 323203), PE-anti-human-IgGFC (purchased from Biolegend, article number 409311), PE-anti-hunman-CD14 (purchased from Biolegend, article number 367103), PE-anti-hunman-CD34 (purchased from Biolegend, article number 343505), PE-anti-hunman-CD31 (purchased from Biolegend, catalog number 303105) 5ul;

[0044] 2.3 Add the antibody to the cell suspension added to the different tubes in step 2.1, incubate for 15 minutes...

Example Embodiment

[0047] Example 3 Extraction of human umbilical cord blood mesenchymal stem cell content

[0048] 3.1 Extract stem cell content from human umbilical cord blood mesenchymal stem cell culture medium;

[0049] 3.1.1 Thaw the culture medium stored in the -20°C refrigerator collected during the cell culture process in the 4°C refrigerator;

[0050] 3.1.2 Use a low-temperature centrifuge to control the temperature. When the temperature of the low-temperature centrifuge reaches 4 degrees, transfer the thawed cell culture medium into a 50ml centrifuge tube, centrifuge at 1000r / min for 5min, and filter the supernatant with a 0.45uM filter.

[0051] 3.1.3 Transfer the filtered supernatant to a new 50ml high-speed centrifuge tube and centrifuge at 10000r / min for 20min. The supernatant liquid was discarded, the sediment at the bottom of the centrifuge tube was resuspended with 2% by volume sterile water, and filtered with a 0.45uM filter to obtain the secretory content of human umbilical cord blo...

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 human umbilical cord blood mesenchymal stem cell extract and a preparation method and application thereof. The human umbilical cord blood mesenchymal stem cell extract is prepared in the steps that 1, human umbilical cord blood mesenchymal stem cells are separated from human umbilical cord blood and cultured in a culture medium added with vitamin C; 2, the secreted content of the human umbilical cord blood mesenchymal stem cells is obtained from the culture medium where the human umbilical cord blood mesenchymal stem cells are cultured; 3, the cultured human umbilicalcord blood mesenchymal stem cells are crushed to obtain the non-secreted content of the human umbilical cord blood mesenchymal stem cells is obtained; 4, the secreted content and the non-secreted content are mixed, and the human umbilical cord blood mesenchymal stem cells separated from the human umbilical cord blood are fully used for preparing the extract containing the secreted content and thenon-secreted content. Besides, the invention further discloses a preparation method of the human umbilical cord blood mesenchymal stem cell extract and application of the human umbilical cord blood mesenchymal stem cell extract in cosmetics.

Description

technical field [0001] The invention relates to the technical field of biomedical stem cells and tissue engineering, in particular to an extract of human umbilical cord blood mesenchymal stem cells and its preparation method and application. Background technique [0002] Mesenchymal stem cells (MSCs) are adult stem cells with self-renewal and multi-lineage differentiation potential, present in bone marrow, adipose tissue, umbilical cord blood and various fetal tissues. It can secrete a variety of cytokines and growth factors to promote the proliferation and differentiation of hematopoietic stem cells (HSC). MSCs also have immunomodulatory, anti-inflammatory and tissue repairing effects, which can alleviate graft-versus-host disease (GVHD) and other transplant-related complications. Mesenchymal Stem Cells (MSCs) are currently mainly extracted from spinal cord and umbilical cord blood. Cord blood is the blood that remains in the placenta and umbilical cord after the baby is ...

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): C12N5/0775A61K8/99A61K8/98A61Q19/08
CPCA61K8/983A61K8/99A61Q19/08C12N5/0665C12N2500/02C12N2500/38
Inventor 艾立平蒋明贵颜丽刘阳
Owner 湖南艾佳生物科技股份有限公司
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