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Culture method of autologous peripheral blood stem cells for treating degenerative arthritis

A technology for peripheral blood stem cells and degenerative arthritis, applied in the field of autologous peripheral blood stem cell culture, can solve the problems of high price, small number of peripheral blood hematopoietic stem cells, limited sources of peripheral blood hematopoietic stem cells, etc., to improve digestion speed, avoid Effect of virus or mycoplasma contamination, reduction of insoluble tissue pieces

Pending Publication Date: 2021-04-27
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] The technical problems to be solved by the present invention: the problem of limited source of peripheral blood hematopoietic stem cells and high price; the problem of small number of peripheral blood hematopoietic stem cells

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] A method for culturing autologous peripheral blood stem cells for treating degenerative arthritis, comprising the following steps:

[0027] Step 1: Treatment of adipose tissue: Human adipose tissue is collected by fat excision or liposuction under local anesthesia, and placed in sterile saline containing antibiotics and antifungal drugs prepared in advance. Wash and remove visible vascular tissue and remove most of the blood cells, centrifuge to remove excess aqueous solution and blood, then add phosphate buffered saline (PBS), transfer to 37°C air bath incubator, vortex at 100r / min, and repeat rinsing. Each time for 30 minutes, use a pipette to suck out the liquid in the lower layer of the tissue until the sucked out liquid is clear;

[0028] Step 2: Isolation of adipose-derived mesenchymal stem cells: add an equal volume of digestive solution to the obtained product in step 1 for digestion, put it in a shaker, rotate and digest at 150r / min at 37°C for 30min, add serum...

Embodiment 2

[0033] A method for culturing autologous peripheral blood stem cells for treating degenerative arthritis, comprising the following steps:

[0034] Step 1: Treatment of adipose tissue: Human adipose tissue is collected by fat excision or liposuction under local anesthesia, and placed in sterile saline containing antibiotics and antifungal drugs prepared in advance. Wash and remove visible vascular tissue and remove most of the blood cells, centrifuge to remove excess aqueous solution and blood, then add phosphate buffered saline (PBS), transfer to 37°C air bath incubator, vortex at 100r / min, and repeat rinsing. Each time for 30 minutes, use a pipette to suck out the liquid in the lower layer of the tissue until the sucked out liquid is clear;

[0035] Step 2: Separation of adipose-derived mesenchymal stem cells: add an equal volume of digestive solution to the obtained product in step 1 for digestion, put it in a shaker, rotate and digest at 150r / min at 37°C for 30-50min, then ...

Embodiment 3

[0040] A method for culturing autologous peripheral blood stem cells for treating degenerative arthritis, comprising the following steps:

[0041] Step 1: Treatment of adipose tissue: Human adipose tissue is collected by fat excision or liposuction under local anesthesia, and placed in sterile saline containing antibiotics and antifungal drugs prepared in advance. Wash and remove visible vascular tissue and remove most of the blood cells, centrifuge to remove excess aqueous solution and blood, then add phosphate buffered saline (PBS), transfer to 37°C air bath incubator, vortex at 100r / min, and repeat rinsing. Each time for 30 minutes, use a pipette to suck out the liquid in the lower layer of the tissue until the sucked out liquid is clear;

[0042] Step 2: Separation of adipose-derived mesenchymal stem cells: add an equal volume of digestive solution to the obtained product in step 1 for digestion, put it in a shaker, rotate and digest at 150r / min at 37°C for 30-50min, then ...

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Abstract

The invention provides a culture method of autologous peripheral blood stem cells for treating degenerative arthritis. The culture method comprises the following steps of 1, treating adipose tissues; 2, separating the adipose tissue-derived stromal cells; 3, culturing the adipose tissue-derived stromal cells; 4, obtaining an extract of the adipose tissue-derived stromal cells; and 5, culturing the autologous peripheral blood stem cells. The autologous adipose tissue-derived stromal cells are extracted and cultured, and a culture medium formed by culturing and mixing multiple generations of adipose tissue-derived stromal cells is used for culturing the autologous peripheral blood stem cells, so that the problems that peripheral blood hematopoietic stem cells are limited in source and small in number are solved; in addition, the whole extraction method of the autologous adipose tissue-derived stromal cells is optimized, so that the purity of the obtained stem cells is higher; I-type collagenase and IV-type collagenase are utilized, so that the digestion speed is increased; and meanwhile, a serum culture solution is utilized, so that potential virus or mycoplasma pollution is avoided.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for culturing autologous peripheral blood stem cells for treating degenerative arthritis. Background technique [0002] Degenerative arthritis is called osteoarthritis, degenerative arthritis, senile arthritis, and hypertrophic arthritis. It is a degenerative disease caused by aging, obesity, strain, trauma, congenital abnormalities of joints, joint deformities, etc. Degeneration of articular cartilage, reactive hyperplasia of joint margins and subchondral bone caused by many factors. According to research and research, in my country, there are more than 100 million patients with osteoarthritis, accounting for nearly 10% of the total population of my country, and the most common type of osteoarthritis is knee arthritis. According to relevant data, , the disability rate of knee arthritis is the second in the world, and the incidence rate will increase with age. It...

Claims

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Application Information

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IPC IPC(8): C12N5/0789C12N5/0775A61P19/02A61P29/00
CPCC12N5/0647A61P19/02A61P29/00C12N2502/1382C12N2509/00C12N2500/90C12N2500/44C12N2500/32C12N2501/115C12N2501/235C12N2501/11C12N2501/998
Inventor 占震锋李庆静
Owner 上海南滨江细胞生物科技有限公司