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CMM culture medium and application thereof

A medium, separation and culture technology, applied in the field of animal medicine and biology, can solve the problems of replicative aging, loss of stemness of mesenchymal stem cells, hindering stem cell treatment, etc., to improve lameness and pain, strong stem cell activity and proliferation ability , Improve the effect of repairing joint function

Pending Publication Date: 2022-05-24
江苏银丰生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The mesenchymal stem cells obtained by the general culture method widely used at present often have stemness loss and severe replicative senescence, which hinders the effect of stem cell therapy research

Method used

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  • CMM culture medium and application thereof
  • CMM culture medium and application thereof
  • CMM culture medium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Preparation of canine platelet-rich plasma

[0029] The canine platelet-rich plasma is obtained by the following method: through the canine vein, take 5-50 ml of whole blood, add heparin for anticoagulation, centrifuge at 1500rpm for 15min, the centrifuge is divided into three layers, the upper layer is serum, the middle layer is platelet-rich plasma The layer and the lower layer are the red blood cell layer; the plasma layer of the middle layer is extracted under aseptic conditions to obtain canine platelet-rich plasma.

Embodiment 2

[0030] Embodiment 2CMM medium preparation

[0031] The specific concentration of each component of the culture medium is adjusted to prepare a CMM-0 complete culture medium, which includes DMEM / F12 culture medium and the canine platelet-rich plasma described in Example 1 that accounts for 5% of the DMEM / F12 culture medium by volume, and also includes 15μg / L selenium, 70mg / LL-ascorbic acid, 200μg / L hydrocortisone, 20mg / L heparin, 500mg / LNaHCO 3 and 400mg / L Transferrin.

Embodiment 3

[0032] Embodiment 3CMM medium preparation

[0033] The specific concentration of each component of the medium was adjusted to prepare a CMM-1 complete medium, which included DMEM / F12 medium and dog platelet-rich plasma with a volume percentage of 1% in the DMEM / F12 medium, and also included 10 μg / L selenium Elements, 55mg / L L-ascorbic acid, 200μg / L hydrocortisone, 15mg / L heparin, 450mg / L NaHCO 3 and 250mg / L Transferrin.

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PUM

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Abstract

The invention discloses a CMM culture medium and application thereof, the CMM culture medium comprises a DMEM / F12 culture medium and dog platelet-rich plasma accounting for 1-10% of the volume of the DMEM / F12 culture medium, and also comprises 10-20 [mu] g / L of selenium element, 55-75 mg / L of L-ascorbic acid, 200-500 [mu] g / L of hydrocortisone, 15-30 mg / L of heparin, 450-500 mg / L of NaHCO3 and 250-400 mg / L of LTransferrin. The invention also discloses an application of the CMM culture medium in isolated culture of canine umbilical cord blood mesenchymal stem cells. Compared with umbilical cord mesenchymal stem cells obtained by a general culture method, the canine umbilical cord mesenchymal stem cells obtained by the separation culture method have higher stem cell activity and value-added ability. The obtained canine umbilical cord mesenchymal stem cells have an obvious improvement effect on canine arthritis.

Description

technical field [0001] The invention relates to a complete culture medium for mesenchymal stem cells and applications thereof, in particular to a complete culture medium for canine umbilical cord mesenchymal stem cells and applications thereof, belonging to the fields of animal medicine and biology. Background technique [0002] The application of stem cells in regenerative medicine and tissue engineering is rapidly emerging in both veterinary experiments and clinical practice. On the one hand, preclinical efficacy and safety testing of new human stem cell therapies must be evaluated using two animal species, namely first rodent species (preferably rats) and second non-rodent larger animals more similar to humans . Mesenchymal stem cells (MSCs) are generally considered superior to rodents in the study of mesenchymal stem cells (MSCs) for therapeutic applications in human disease. Compared with rodents, dogs are more similar than rodents to humans in terms of anatomy, patho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61K35/28A61P19/02A61P29/00
CPCC12N5/0668A61K35/28A61P19/02A61P29/00C12N2509/10C12N2509/00C12N2500/05C12N2500/38C12N2501/39C12N2501/91C12N2500/12C12N2500/24
Inventor 袁术斌袁荃樊德俊孙博
Owner 江苏银丰生物工程有限公司
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