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Cryopreservation resuscitation solution for sweat gland-like cells and cryopreservation and resuscitation method for maintaining cell activity

A technology of sweat gland-like cells and resuscitation fluid, applied in the field of human sweat gland regeneration research, can solve the problems of low survival efficiency and unfavorable resuscitation applications, and achieve the effects of clear components, high survival rate, and easy use.

Active Publication Date: 2016-04-20
FIRST HOSPITAL AFFILIATED TO GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the survival rate of sweat gland-like cells after cryopreservation and recovery after induction is low, only about 30%, which is not conducive to the application of recovery

Method used

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  • Cryopreservation resuscitation solution for sweat gland-like cells and cryopreservation and resuscitation method for maintaining cell activity
  • Cryopreservation resuscitation solution for sweat gland-like cells and cryopreservation and resuscitation method for maintaining cell activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] (1) Cell cryopreservation

[0035] Preparation of cryopreservation solution: The components of the cryopreservation solution used in this example are: the base fluid serum substitute contains:

[0036] Human epidermal growth factor at a concentration of 70 ng / mL, recombinant human keratinocyte growth factor-II at a concentration of 40 ng / mL, triiodothyronine at a concentration of 25 ng / mL, hydrocortisone hemisuccinate at a concentration of 0.7 μg / mL, insulin-transferrin-sodium selenite, the concentration is 0.05mL / mL, bovine serum albumin, the concentration is 0.4g / mL, and the prepared cryopreservation solution is pre-cooled in a refrigerator at 4°C for 30 minutes .

[0037]Take the induced sweat gland-like cells, digest them, collect them and centrifuge them, then add the pre-cooled cryopreservation solution so that the cell concentration is 0.5-2×10 6 / mL, transfer to cryopreservation tubes at 1.5mL / branch, and pre-cool at 4°C for 30 minutes. Put it into the temper...

Embodiment 2

[0047] (1) Cell cryopreservation

[0048] The composition of the cryopreservation solution used in this example is: the base solution Knock-out serum substitute contains:

[0049] Human epidermal growth factor at a concentration of 80 ng / mL, recombinant human keratinocyte growth factor-II at a concentration of 50 ng / mL, triiodothyronine at a concentration of 30 ng / mL, hydrocortisone hemisuccinate at a concentration of 0.8 μg / mL, insulin-transferrin-sodium selenite, the concentration is 0.1mL / mL, bovine serum albumin, the concentration is 0.6g / mL.

[0050] The cryopreservation operation is the same as in Example 1.

[0051] (2) Cell recovery

[0052] The components of the resuscitation fluid used in this example are: the base fluid Knock-out serum substitute contains: human epidermal growth factor at a concentration of 80 ng / mL, recombinant human keratinocyte growth factor-II at a concentration of 50 ng / mL, triiodine Thyronine at a concentration of 30 ng / mL, hydrocortisone h...

Embodiment 3

[0057] (1) Cell cryopreservation

[0058] The composition of the cryopreservation solution used in this example is: the base solution Knock-out serum substitute contains:

[0059] Human epidermal growth factor at a concentration of 75 ng / mL, recombinant human keratinocyte growth factor-II at a concentration of 45 ng / mL, triiodothyronine at a concentration of 27 ng / mL, hydrocortisone hemisuccinate at a concentration of 0.75 μg / mL, insulin-transferrin-sodium selenite, the concentration is 0.07mL / mL, bovine serum albumin, the concentration is 0.4g / mL.

[0060] The cryopreservation operation is the same as in Example 1.

[0061] (2) Cell recovery

[0062] The composition of the resuscitation fluid used in this embodiment is: the base fluid Knock-out serum substitute contains:

[0063] Human epidermal growth factor at a concentration of 75 ng / mL, recombinant human keratinocyte growth factor-II at a concentration of 45 ng / mL, triiodothyronine at a concentration of 27 ng / mL, hydro...

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Abstract

The invention provides sweat gland cell cryopreservation liquid and recovery liquid. The following components are added into a serum substitution to prepare the liquid, wherein the components comprises a human epidermal growth factor whose concentration is 50 to100ng / mL, a recombinant human keratinocyte growth factor-II whose concentration is 10 to 100ng / mL, triiodothyronine whose concentration is 10 to 50ng / mL, hydrocortisone hemisuccinate whose concentration is 0.5 to 1 microgramme / mL, insulin-transferrin-sodium selenite whose concentration is 0.01 to 0.2mL / mL, and bovine serum albumin whose concentration is 0.01 to 1g / mL. The invention further provides a cell cryopreservation recovery method using the cryopreservation liquid and the recovery liquid. Compared with the traditional cryopreservation recovery method, the method provided by the invention has the advantage of high cryopreservation recovery survival rate, does not affect the characteristics of a sweat gland cell, does not change the differentiation state of the sweat gland cell, and solves the problem of difficult sweat gland cell storage recovery.

Description

technical field [0001] The present invention relates to the research field of human sweat gland regeneration, and more specifically, relates to a cryopreservation and resuscitation method for maintaining activity of sweat gland-like cells, and a cryopreservation solution and a resuscitation solution used in the method. Background technique [0002] The skin is the largest organ in the human body and can remove 25% of the body's heat to keep the body temperature relatively constant. Patients with extensive deep burns lose their ability to secrete sweat due to the destruction of sweat glands or the blockage of their secretion ducts by scar tissue, and the dysfunction of skin temperature regulation seriously affects the quality of life of patients. Therefore, it is of great significance to restore the perspiration function of patients with extensive deep burns to improve the quality of life of patients. The artificial skin developed at present has no skin appendages such as sw...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02C12N5/071
Inventor 付小兵马奎黄沙盛志勇
Owner FIRST HOSPITAL AFFILIATED TO GENERAL HOSPITAL OF PLA
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