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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
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  • 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 cells is single, the present invention proposes an extract of human umbilical cord blood mesenchymal stem cells and its preparation method and application

Method used

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  • 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

Embodiment 1

[0035] Example 1 Isolation and cultivation 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 long-term cooperation with our company, select healthy cesarean section women under the age of 30, without genetic diseases, without infectious diseases, and with negative anti-HIV and hepatitis B serum markers to collect cord blood . After the collection is completed, the cord blood is placed in an ice box and immediately transported to the laboratory for separation and processing. Among them, the involved mothers were informed and agreed to the isolated cord blood experiment protocol.

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

[0039] The collected umbilical cord blood was diluted with phosphate buffered saline (referred to as PBS) to an equal volume, and then density gradient centrifugation was performed using lymph separati...

Embodiment 2

[0041] Example 2 Cell Type Identification of Isolated Human Umbilical Cord Blood Mesenchymal Stem Cells

[0042]2.1 The umbilical cord blood mesenchymal stem cells cultured in vitro to the third generation were taken, digested, centrifuged, and the cell density was adjusted to 1x106 cells / ml with PBS, and distributed into multiple tubes.

[0043] 2.2 Take the following antibodies respectively: ITC-anti-human-IgG FC (purchased from Biolegend, product number 409309), FITC-anti-hunman-CD44 (purchased from Biolegend, product number 338803), FITC-anti-hunman-CD105 (purchased from Biolegend , Cat. No. 323203), PE-anti-human-IgGFC (purchased from Biolegend, Cat. No. 409311), PE-anti-hunman-CD14 (purchased from Biolegend, Cat. 343505), PE-anti-hunman-CD31 (purchased from Biolegend, item number 303105) 5ul;

[0044] 2.3 Add the antibody to the cell suspension added to the different tubes in step 2.1, incubate at room temperature for 15 minutes in the dark, and replenish the volume of ...

Embodiment 3

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

[0048] 3.1 extract the stem cell content in the human umbilical cord blood mesenchymal stem cell culture medium;

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

[0050] 3.1.2 Use a cryogenic centrifuge to control the temperature. When the temperature of the cryogenic centrifuge reaches 4°C, transfer the thawed cell culture medium into a 50ml centrifuge tube, and 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. Discard the supernatant, resuspend the sediment at the bottom of the centrifuge tube with 2% volume ratio sterile water, and filter with a 0.45uM filter to obtain the secreted content of human umbilical cord blood mesenchymal stem cells in ...

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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

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Application Information

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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
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