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Method for extracting autologous adipose-derived stem cells for treating degenerative arthritis

A technology for degenerative arthritis and extraction methods, which is applied in the field of stem cell preparation, can solve the problems affecting the induction and formation process of stem cell proliferation, differentiation, cumbersome stem cell process, and low purity, and achieve short proliferation time, strong in vitro expansion ability, and improved survival. rate effect

Inactive Publication Date: 2021-05-14
上海南滨江细胞生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In the prior art, the extraction of adipose-derived stem cells is mostly digested with trypsin. The process of extracting stem cells is cumbersome and the purity is not high, which affects the proliferation and differentiation of stem cells in the later stage and even the induction process. Therefore, a stem cell with high purity, simple method and high cell activity is provided. The extraction method is a technical problem that needs to be solved at present

Method used

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Examples

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Embodiment 1

[0023] A method for extracting autologous adipose-derived stem cells for treating degenerative arthritis, comprising the following steps:

[0024] Step 1. Under aseptic conditions, take normal abdominal subcutaneous fat tissue, remove obvious connective tissue and small blood vessels visible to the naked eye, wash the fat tissue 3 times with PBS buffer, and then place it in PBS buffer to inactivate it. Bacteria ophthalmic scissors shred the adipose tissue, and then use ophthalmic scissors to carefully remove the spasm, fat droplets and fat granules to obtain small pieces of fat;

[0025] Step 2: Add 1mL of serum to a sterile petri dish and shake it evenly, put 25 fat pieces obtained in the first step into a sterile petri dish, keep the interval between the fat pieces at 4mm, and then Add 3 mL of complete culture solution to a sterile petri dish, let it stand for 3 minutes, then slowly invert the petri dish, and place the inverted petri dish in 3% CO 2 In the incubator, after ...

Embodiment 2

[0034] A method for extracting autologous adipose-derived stem cells for treating degenerative arthritis, comprising the following steps:

[0035] Step 1. Under aseptic conditions, take normal abdominal subcutaneous fat tissue, remove obvious connective tissue and small blood vessels visible to the naked eye, wash the fat tissue 4 times with PBS buffer, and then place it in PBS buffer to inactivate it. Bacteria ophthalmic scissors shred the adipose tissue, and then use ophthalmic scissors to carefully remove the spasm, fat droplets and fat granules to obtain small pieces of fat;

[0036] The second step, add 2mL serum to the sterile petri dish and shake it to spread evenly, put 30 fat pieces obtained in the first step into the sterile petri dish, keep the interval between the fat pieces 5mm, and then Add 3 mL of complete culture solution to a sterile petri dish, let it stand for 4 minutes, then slowly invert the petri dish, and place the inverted petri dish in a volume fractio...

Embodiment 3

[0045] A method for extracting autologous adipose-derived stem cells for treating degenerative arthritis, comprising the following steps:

[0046] Step 1. Under aseptic conditions, take normal abdominal subcutaneous fat tissue, remove obvious connective tissue and small blood vessels visible to the naked eye, wash the fat tissue 4 times with PBS buffer, and then place it in PBS buffer to inactivate it. Bacteria ophthalmic scissors shred the adipose tissue, and then use ophthalmic scissors to carefully remove the spasm, fat droplets and fat granules to obtain small pieces of fat;

[0047] In the second step, add 2.5mL serum to the sterile petri dish and shake it evenly, put 28 pieces of fat pieces obtained in the first step into the sterile petri dish, keep the interval between the fat pieces at 5mm, Then add 3.5mL of complete culture solution to the sterile Petri dish, let it stand for 4 minutes, then slowly invert the Petri dish, and place the inverted Petri dish in a volume ...

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Abstract

The invention discloses a method for extracting autologous adipose-derived stem cells for treating degenerative arthritis, which comprises the following steps: firstly, removing connective tissues, small blood vessels, fat droplets and fat granules from normal abdominal subcutaneous adipose tissues under a sterile condition to obtain small adipose blocks; then culturing the adipose cubes in a complete culture solution, carrying out trypsin digestion and centrifugal treatment to obtain adipose-derived stem cells, and finally, subculturing the adipose-derived stem cells in the complete culture solution and an anti-apoptosis protective agent which has the functions of promoting the proliferation and differentiation of animal hematopoietic stem cells and also has the functions of achieving anti-apoptosis protection effect in adipose-derived stem cell culture process, improving the survival rate of adipose-derived stem cells and the later induced differentiation success rate.The tissue block culture method adopted by the invention is simple to operate, strong in-vitro amplification capability and short in proliferation time, has multidirectional differentiation potential, capability of forming cartilage cells through induced differentiation culture, and can treat degenerative arthritis.

Description

technical field [0001] The invention belongs to the technical field of stem cell preparation, and in particular relates to a method for extracting autologous adipose-derived stem cells for treating degenerative arthritis. Background technique [0002] Degenerative arthritis is also called osteoarthritis, senile arthritis, and hypertrophic arthritis. The main symptom is joint pain, which is often rest pain. Another symptom is joint stiffness, which often occurs when getting up in the morning or after the joints maintain a certain position for a long time during the day. Examination of the affected joints shows swelling and tenderness of the joints, and friction or "clicking" sounds during activities. In severe cases There may be muscle wasting and joint deformities. [0003] Adipose-derived stem cells are a type of stem cells that can self-renew and reproduce. They have the multi-directional differentiation potential of general stem cells and the stable multi-metabolism valu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61P19/02A61P29/00
CPCA61P19/02A61P29/00C12N5/0667C12N2500/34C12N2500/84C12N2509/00
Inventor 占震锋李庆静
Owner 上海南滨江细胞生物科技有限公司
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