A kind of mesenchymal stem cell culture medium

A technology of basal medium and serum-free medium, applied in cell culture active agents, tissue culture, animal cells, etc., can solve the problems of insufficient antibody inhibitors and insufficient types of Rho kinase inhibitors, etc., and achieve the purpose of inhibiting invasion and migration ability, good application value, and the effect of promoting proliferation

Active Publication Date: 2022-07-05
诺赛联合(北京)生物医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But at present, there are not enough types of Rho kinase inhibitors,...

Method used

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  • A kind of mesenchymal stem cell culture medium
  • A kind of mesenchymal stem cell culture medium
  • A kind of mesenchymal stem cell culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Preparation of Rho GTPase monoclonal antibody

[0028] Rho GTPase 1 (RND1) was used as the immunogen (Shanghai Hewu Biotechnology Co., Ltd. purchased in May 2020).

[0029] Using Rho GTPase 1 as the immunogen, with 5-fold dilution of Montanide TMGEL 01 ST polymer adjuvant was mixed with equal volume, and BALB / c mice were immunized by subcutaneous multi-point injection, 3 times in total, each immunization interval was 2 weeks, and the dose was 100 μg / mice. Seven days after the third immunization, the mice were immunized with the immunogen without adjuvant. Seven days after each immunization, blood was collected to detect the antibody level by indirect ELISA. After 4 days, the mouse splenocytes with the highest titer were taken for fusion.

[0030] SP2 / 0 cells were fused with splenocytes under the action of PEG1500, and after fusion, DMEM medium (DMEM dry powder 13.5 g / L, NaHCO) was used. 3 3.7g / L, L-glutamine 0.2g / L) was added to HAT and HT for liquid excha...

Embodiment 2

[0039] Example 2 Preparation of serum-free medium for stem cell proliferation-promoting

[0040] Described serum-free culture medium is made up of basal medium and the supplementary component added in basal medium, described basal medium is DMEM / F12 medium, and described supplementary component is L-glutamine, non-essential amino acid, L-glutamine. -Ascorbic acid, sodium selenite, fibronectin, ethanolamine, hydrocortisone, trypsin inhibitor, human transferrin, human insulin, bFGF, TGF-β1, PDGF-BB, coenzyme A, sodium pyruvate, and EGF And RND1-5 monoclonal antibody; wherein, the concentration of L-glutamine is 2mM, the concentration of non-essential amino acid is 2mM, the concentration of L-ascorbic acid is 50mg / L, the concentration of sodium selenite is 10μg / L, the concentration of fiber The concentration of zonulin was 20 mg / L, the concentration of ethanolamine was 2 mg / L, the concentration of hydrocortisone was 10 mg / L, the concentration of trypsin inhibitor was 1 mg / L, the ...

Embodiment 3

[0042] Example 3 Preparation of serum-free medium control group for stem cell proliferation-promoting

[0043] Described serum-free culture medium is made up of basal medium and the supplementary component added in basal medium, described basal medium is DMEM / F12 medium, and described supplementary component is L-glutamine, non-essential amino acid, L-glutamine. -Ascorbic acid, sodium selenite, fibronectin, ethanolamine, hydrocortisone, trypsin inhibitor, human transferrin, human insulin, bFGF, TGF-β1, PDGF-BB, coenzyme A, sodium pyruvate, and EGF ; Among them, the concentration of L-glutamine is 2mM, the concentration of non-essential amino acids is 2mM, the concentration of L-ascorbic acid is 50mg / L, the concentration of sodium selenite is 10μg / L, the concentration of fibronectin is 20mg / L L, the concentration of ethanolamine is 2mg / L, the concentration of hydrocortisone is 10mg / L, the concentration of trypsin inhibitor is 1mg / L, the concentration of human transferrin is 10m...

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Abstract

The present invention relates to a medium for mesenchymal stem cells. The present invention provides a monoclonal antibody against Rho GTPase, the antibody can specifically inhibit the activity of Rho GTPase, thereby regulating the expression of downstream ROCK1 protein, the antibody can promote the proliferation of stem cells, and can also inhibit cancer The ability of cell invasion and migration has good application value.

Description

technical field [0001] The invention relates to the biological field, and more particularly to a mesenchymal stem cell culture medium. Background technique [0002] Rho GTPase (Rho GTPase) belongs to the Ras superfamily and is involved in cell migration, phagocytosis, contraction and adhesion. ROCK, also known as Rho kinase (Rho-associated kinase), is currently the most detailed Rho downstream target effector molecule. The Rho / ROCK signaling pathway induces cytoskeletal reorganization, cell migration, and stress fiber formation, and is associated with various physiological functions such as endothelial permeability, tissue contraction and growth, and is involved in diabetes, eye diseases, tumors, heart disease, nerve injury diseases, The occurrence of diseases such as hypertension, radiation injury and leukemia has attracted more and more attention as a drug development target. [0003] The Rho / ROCK signaling pathway activates downstream ROCK through the binding of Rho to ...

Claims

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Application Information

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IPC IPC(8): C07K16/40C12N5/0775A61K39/395A61P35/00
CPCC07K16/40C12N5/0667A61P35/00C07K2317/56C12N2501/73
Inventor 亓爱杰李少波陈清轩李斌斌
Owner 诺赛联合(北京)生物医学科技有限公司
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