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Low-temperature preservation and transportation culture medium

A technology of cryopreservation and culture medium, applied in the culture process, tissue culture, cell culture active agent, etc., can solve the problems of reduced tissue viability, inability to cultivate for a long time, loss of pigment-induced response, etc., to increase the range, facilitate long-distance transportation, The effect of reducing the damage of cryopreservation

Active Publication Date: 2020-05-05
GUANGDONG BOXI BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The object of the present invention is to provide a melanocyte-containing skin model for the problems of reduced tissue viability, loss of pigment-induced response, and inability to be cultured for a long time after storage in the prior art during storage and transportation of the skin model containing melanocytes. Cryopreserved Transport Medium for Cryopreservation

Method used

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  • Low-temperature preservation and transportation culture medium
  • Low-temperature preservation and transportation culture medium
  • Low-temperature preservation and transportation culture medium

Examples

Experimental program
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Effect test

Embodiment 1

[0054] This embodiment provides a melanin skin model culture solution, which includes basal culture solution, hydrocortisone, insulin, β-fibroblast growth factor, epidermal growth factor and keratinocyte growth factor, Epinephrine, phorbol 12-myristate-13-acetate, stem cell factor, glutamine, calcium chloride, and antibiotics. The basal culture medium is K-SFM or F12 / DMEM, and the volume ratio of F12 and DMEM in the F12 / DMEM is 1:1.

[0055] The preparation method of the melanin skin model culture fluid described in the present embodiment comprises the following steps:

[0056] 1) Preparation of melanin skin model culture medium: Based on the commercially available epidermal cell culture medium K-SFM, take 100ml of K-SFM and add 50 μg / L hydrocortisone and 10 μg / L insulin to it at the indicated final concentration , β-fibroblast growth factor 2μg / L, epidermal growth factor 2ng / L, the final concentration of keratinocyte growth factor in the culture medium is 5ng / L, epinephrine ...

Embodiment 2

[0058] This embodiment provides a low-temperature storage transport medium, which includes the following components: the melanin skin model culture solution described in Example 1, 4-hydroxyethylpiperazineethanesulfonic acid, agarose , sucrose, glycine, adenosine, alpha-tocopherol, vitamin C, sphingosine-1-phosphate, Y-27632 and heparin.

[0059] The low-temperature preservation transportation medium described in this embodiment is prepared, and the preparation method is as follows:

[0060] Solution A: Take 100ml melanin model culture solution, add 4-hydroxyethylpiperazineethanesulfonic acid 20mM / L, sucrose 2mM / L, glycine 25mM / L, adenosine 0.2mM / L , α-tocopherol 0.05mM / L, vitamin C 0.05mM / L, sphingosine-1-phosphate 10μM / L, Y-27632 10μM, heparin 3μg / L, after fully dissolved, use a 0.22μm membrane filter to sterilize .

[0061] Liquid B: Weigh 2 g of agarose, add 100 ml of deionized water to dissolve it, and make an agarose solution with a concentration of 2%, and sterilize i...

Embodiment 3

[0067] This embodiment provides a melanin skin model culture solution, which is prepared by the following method: take 100ml of F12 / DMEM (1:1) culture medium, add hydrogenated Cortisone 50 μg / L, insulin 10 μg / L, β-fibroblast growth factor 2 μg / L, epidermal growth factor 2 ng / L, the final concentration of keratinocyte growth factor in the culture medium was 5 ng / L, epinephrine 50 μg / L L, 12-myristic acid-13-acetic acid phorbol 12ng / L, stem cell factor 10μg / L, glutamine 6mM / L, calcium chloride 1.5mM / L, 1% penicillin and streptomycin.

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Abstract

The invention provides a low-temperature preservation and transportation culture medium. The low-temperature preservation and transportation culture medium comprises the following components: a melanin skin model culture solution, a buffer system, agarose, sucrose, glycine, adenosine, alpha-tocopherol, vitamin C, sphingosine-1-phosphoric acid, Y-27632 and heparin, wherein the melanin skin model culture solution comprises a basic culture solution, hydrocortisone, insulin, growth factors, epinephrine, 12-myristic acid-13-phorbol acetate, stem cell factors, glutamine, calcium chloride and antibiotics.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering, and in particular relates to a low-temperature preservation transport medium. Background technique [0002] Melanocytes are derived from the ectodermal neural crest and widely exist in various tissues of the body, such as epidermis, hair follicles, dermis, eyes, peripheral nerves, sympathetic nerve trunks, pia mater and other structures. Melanocytes produce melanin granules, which are transmitted to the surrounding keratinocytes and redistributed in the keratinocytes. Melanin granules can absorb ultraviolet rays and protect the skin from damage caused by ultraviolet rays. The in vitro culture of melanocytes and the construction of 3D skin models are of great significance for the study of skin pigment metabolism, skin aging, and pigment disorders. In the past ten years, the method of in vitro culture of melanocytes has gradually matured, from a single melanocyte culture to a skin model...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071A01N1/02
CPCC12N5/0629A01N1/0215A01N1/0226A01N1/0221C12N2501/39C12N2501/33C12N2501/115C12N2501/11C12N2501/117C12N2501/81C12N2501/999C12N2501/125C12N2500/32C12N2500/14
Inventor 李润芝卢永波张勇杰
Owner GUANGDONG BOXI BIO TECH CO LTD
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