Adipose mesenchymal stem cell preparation kit

A adipose stem cell and fat technology, which is applied in the field of kits for preparing adipose-derived mesenchymal stem cells, can solve the problems of slow progress in scientific research, and achieve the effects of reducing infection risk, convenient use and low cost.

Inactive Publication Date: 2015-09-23
青岛瑞思科生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is urgent to develop a simple, rapid, high-tech, and safe method for obtaining a large amount of adipose-derived mesenchymal stem cells. However, there is no kit for preparing adipose-derived mesenchymal stem cells in the existing technology, which makes the progress of related scientific research work slow.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 Preparation and application of a kit for preparing adipose-derived mesenchymal stem cells

[0018] 1. Preparation of a kit for preparing adipose-derived mesenchymal stem cells

[0019] 1. Solution configuration

[0020] The kit for preparing adipose-derived mesenchymal stem cells of the present invention includes the following reagents:

[0021] (1) Preparation method of fat preservation solution: penicillin (brand Amresco, product number 0242) 200mg, streptomycin (brand Amresco, product number 0382) 200mg, amphotericin (brand Amresco, product number E437) 5mg, dissolved in 1000ml DMEM medium 0.22um filter sterilization is sufficient.

[0022] (2) Preparation method of fat washing solution: dissolve 200mg of penicillin, 200mg of streptomycin, and 5mg of amphotericin in 1000ml of normal saline and sterilize by 0.22um filter.

[0023] (3) Preparation method of lipolysis solution: Dissolve 1 g of collagenase I (brand sigma, product number C0130) in 1000 ml ...

Embodiment 2

[0041] Example 2 Preparation and application of a kit for preparing adipose-derived mesenchymal stem cells

[0042] 1. Preparation of a kit for preparing adipose-derived mesenchymal stem cells

[0043] 1. Solution configuration

[0044] The kit for preparing adipose-derived mesenchymal stem cells of the present invention includes the following reagents:

[0045] (1) Preparation method of fat preservation solution: dissolve 100mg of penicillin, 100mg of streptomycin, and 2.5mg of amphotericin in 1000ml of DMEM / F12 medium, filter and sterilize at 0.22um, and dispense them into aliquots.

[0046] (2) Preparation method of fat washing solution: 100mg of penicillin, 100mg of streptomycin, 2.5mg of amphotericin, dissolved in 1000ml of normal saline 0.22um, filtered and sterilized for subpackaging.

[0047] (3) Preparation method of lipolysis liquid: Dissolve 2 g of collagenase I in 1000 ml of DMEM / F12 medium, and filter and sterilize with a 0.22um filter.

[0048] (4) Cell wa...

Embodiment 3

[0057] Example 3 Preparation and application of a kit for preparing adipose-derived mesenchymal stem cells

[0058] 1. Preparation of a kit for preparing adipose-derived mesenchymal stem cells

[0059] 1. Solution configuration

[0060] The kit for preparing adipose-derived mesenchymal stem cells of the present invention includes the following reagents:

[0061] (1) Configuration method of fat preservation solution: dissolve 150mg of penicillin, 150mg of streptomycin, and 2.5mg of amphotericin in 1000ml of DMEM medium, filter and sterilize at 0.22um, and then dispense them.

[0062] (2) Preparation method of fat washing solution: dissolve 150mg of penicillin, 150mg of streptomycin, and 2.5mg of amphotericin in 1000ml of normal saline and filter to sterilize at 0.22um.

[0063] (3) Preparation method of lipolysis solution: Dissolve 2 g of collagenase I in 1000 ml of DMEM medium, and filter and sterilize with a 0.22um filter.

[0064] (4) Cell washing solution configurati...

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PUM

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Abstract

The present invention discloses an adipose mesenchymal stem cell preparation kit, the kit includes a fat storage solution, a fat washing liquid, a fat decomposition liquid, a cell washing liquid, a cell culture liquid, a cell digestion liquid and a cell cryopreservation liquid; the fat storage solution is a DMEM, DMEM / F12, MEM or RPMI-1640 medium including 100-200ug / ml of penicillin, 100-200ug / ml of streptomycin and 2.5-5 mug / ml of amphotericin; the fat washing liquid is normal saline, PBS, D-Hanks or HBS including 100-200ug / ml of penicillin, 100-200ug / ml of streptomycin and 2.5-5mug / ml of amphotericin; the fat decomposition liquid is a DMEM, DMEM / F12, MEM or RPMI-1640 medium including 0.75-2mg / ml of collagenase I; the cell washing liquid is PBS, D-Hanks or HBSS; the cell culture liquid is a mesenchymal stem cell serum-free growth medium, the cell digestion liquid is a PBS, D-Hanks or HBSS solution including 1.25-2.5mg / ml of Trypsin-EDTA; and the cell cryopreservation liquid is a mesenchymal stem cell serum-free growth medium including 60-200mg / ml of DMSO.

Description

[0001] technical field [0002] The invention belongs to the technical field of stem cells, and relates to a kit for preparing adipose mesenchymal stem cells. Background technique [0003] In 2001, Zuk et al. discovered that there is a type of cell with multi-directional differentiation potential in adipose tissue, that is, adipose-derived mesenchymal stem cells, referred to as adipose-derived stem cells. Its biological properties are similar to bone marrow mesenchymal stem cells, and can differentiate into various cells such as fat, bone, cartilage, muscle, endothelium, hematopoietic, liver, pancreatic islets and nerves. [0004] Adipose tissue is rich in reserves in the human body, easy to obtain and less traumatic, and has broad application prospects in tissue engineering, organ repair, gene therapy, etc. Therefore, adipose-derived mesenchymal stem cells have become a hot spot in the field of stem cells. In January 2012, Cuepistem, an autologous adipose stem cell dr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 张炳强许评尹桂萍崔冰王彩云史光军
Owner 青岛瑞思科生物科技有限公司
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