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Cryopreservation and resuscitation method for pilose antler stem cells

A technology of stem cells and velvet antler, which is applied in the field of cryopreservation and recovery of velvet antler stem cells, can solve the problems of lack of maintenance factors, low cell viability, single composition, etc., and achieve the effect of maintaining stem cell performance, extending storage time, and ensuring activity

Pending Publication Date: 2022-05-24
玺瑞生命科学(深圳)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing cryopreservation solution is relatively simple in composition, lacks necessary maintenance factors, etc., and the cell viability after recovery is low

Method used

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  • Cryopreservation and resuscitation method for pilose antler stem cells

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Experimental program
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Effect test

Embodiment 1

[0022] as Figure 1 As shown, the present invention proposes a deer antler stem cell cryopreservation and resuscitation method, the method steps comprising:

[0023] S1, the preparation of cryoprotective solution; the configuration ratio of cryoprotective solution is 20%-30% DMSO, 2%-7% human blood albumin, 0.5%-1% trehalose, 10%-12% dextran and 50%-70% physiological sodium chloride solution;

[0024] S2, the deer antler stem cells are mixed with the cryoprotective solution in S1 to obtain the cell suspension, as for the sterile cryovial tube;

[0025] S3, according to the order of 4 °C 1h, -20 °C 1h, -80 °C 1h, -196 °C 1h order for liquid nitrogen freezing;

[0026] S4: Put the cryovial into a 37 °C water bath and shake until the cell suspension is completely melted.

[0027] S5, centrifugation, buffer wash to remove cryoprotectant.

Embodiment 2

[0029] as Figure 1 As shown, the present invention proposes a deer antler stem cell cryopreservation and resuscitation method, the method steps comprising:

[0030] S1, the preparation of cryoprotective solution; the configuration ratio of cryoprotective solution is 20%-30% DMSO, 2%-7% human blood albumin, 0.5%-1% trehalose, 10%-12% dextran and 50%-70% physiological sodium chloride solution;

[0031] S2, the deer antler stem cells are mixed with the cryoprotective solution in S1 to obtain the cell suspension, as for the sterile cryovial tube;

[0032] S3, according to the order of 4 °C 1h, -20 °C 1h, -80 °C 1h, -196 °C 1h order for liquid nitrogen freezing;

[0033] S4: Put the cryovial into a 37 °C water bath and shake until the cell suspension is completely melted.

[0034] S5, centrifugation, buffer wash to remove cryoprotectant.

[0035] Further, the cryoprotective solution is configured in proportion to 20% DMSO, 2% human blood albumin, 0.5% trehalose, 10% dextran and 7.5% ph...

Embodiment 3

[0040] as Figure 1 As shown, the present invention proposes a deer antler stem cell cryopreservation and resuscitation method, the method steps comprising:

[0041]S1, the preparation of cryoprotective solution; the configuration ratio of cryoprotective solution is 20%-30% DMSO, 2%-7% human blood albumin, 0.5%-1% trehalose, 10%-12% dextran and 50%-70% physiological sodium chloride solution;

[0042] S2, the deer antler stem cells are mixed with the cryoprotective solution in S1 to obtain the cell suspension, as for the sterile cryovial tube;

[0043] S3, according to the order of 4 °C 1h, -20 °C 1h, -80 °C 1h, -196 °C 1h order for liquid nitrogen freezing;

[0044] S4: Put the cryovial into a 37 °C water bath and shake until the cell suspension is completely melted.

[0045] S5, centrifugation, buffer wash to remove cryoprotectant.

[0046] Further, the configuration ratio of cryoprotectant is 30% DMSO, 7% human blood albumin, 1% trehalose, 10% dextran and 52% physiological sodium...

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Abstract

The invention relates to the technical field of pilose antler stem cell research, in particular to a pilose antler stem cell cryopreservation and recovery method. The invention relates to a cryopreservation and recovery method for pilose antler stem cells. A cryopreservation solution is prepared from DMSO, human serum albumin, trehalose, dextrose and a physiological sodium chloride solution. Wherein DMSO can reduce gene drift, slow down aging of a cell line and reduce microbial contamination. The human serum albumin has the properties of binding and transporting endogenous and exogenous properties, and is beneficial to keeping the performance of stem cells. The trehalose and the dextrose have a good protection effect on cell viability in the cell freezing, drying and freeze-drying processes. The pilose antler stem cells are subjected to mixing with the cryopreservation liquid, liquid nitrogen freezing and water bath recovery, so that the activity of the pilose antler stem cells is effectively guaranteed, the storage time of the pilose antler stem cells is prolonged, and the study on the pilose antler stem cells is facilitated.

Description

Technical field [0001] The present invention relates to the field of deer antler stem cell research technology, in particular to a deer antler stem cell cryopreservation and resuscitation method. Background [0002] Deer antler velvet is a rare organ in mammals that can be completely regenerated periodically, and is an excellent model for studying organ regeneration in mammals. The periodic regeneration of deer antler velvet is a stem cell-based process, and deer antler stem cells (ASCs) are found in the periosteum (AP) of the raw antler area and the periosteum of the horn stalk (PP). Deer antler stem cells do not chimerize into the entire host tissue like embryonic stem cells, nor disappear completely like mesenchymal stem cells, but are limitedly chimerized into some tissues, especially into the reproductive system. It is of great significance to regenerative medicine. However, the existing cryopreservation solution composition is relatively single, lack of necessary maintenanc...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226A01N1/0284
Inventor 陈丽晶
Owner 玺瑞生命科学(深圳)有限公司
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