Stem cell factor liposome for repairing damaged tissues and preparation method thereof

A stem cell factor, a technology for repairing damage, applied in the field of stem cells, can solve problems such as easy damage and adverse effects of absorption

Inactive Publication Date: 2020-06-12
青岛信迪细胞生物科技开发有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Many of these substances are transported in the form of exosomes and other vesicles, but such membrane structu...

Method used

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  • Stem cell factor liposome for repairing damaged tissues and preparation method thereof
  • Stem cell factor liposome for repairing damaged tissues and preparation method thereof
  • Stem cell factor liposome for repairing damaged tissues and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment 1

[0033] Obtain fresh mesenteric fat from healthy young dogs, then transfer it to a 50ml sterile centrifuge tube, soak and wash it with 2% double-antibody DPBS buffer three times; purify the washed fat tissue, and remove the internal blood vessels and fascia , to obtain high-purity adipose tissue; cut high-purity adipose tissue into 1mm 3 small pieces, and transferred to a 50ml sterile centrifuge tube, added 10ml of 0.25% trypsin digestion solution, 37 ° C shaking digestion for 30 minutes; the digested adipose tissue was centrifuged at 3000r / min for 10 minutes, and the supernatant was taken out. 5 ml MEM culture solution containing 10% FBS, 100 U / ml penicillin and 100 U / ml streptomycin was added to the supernatant taken out to resuspend the pelleted cells to obtain a resuspended cell solution.

[0034] Add an equal volume of 0.4% trypan blue solution to the separated resuspended cell liquid; use the Cellometer Mini automatic cell counter to detect the proportion of viable cells ...

specific Embodiment 2

[0039] The same part of this embodiment and specific embodiment 1 will not be repeated, the difference is

[0040] Weigh 0.4g of soybean lecithin, 0.05g of cholesterol and 0.1g of Tween 80 and dissolve them in 10mL of absolute ethanol, and ultrasonically dissolve them fully; The mixture was introduced into 10 mL of 10% ammonium sulfate solution, and then placed in a round bottom flask; after the solution was evaporated under reduced pressure at 50°C to remove the organic solvent, it was placed in DPBS with pH 7.0 and dialyzed for 24 hours to form liposomes.

[0041] Put the dialyzed liposomes into a water bath shaker at 50°C for oscillation, then slowly add 20ml of the prepared stem cell extract, after the addition is complete, continue to shake at 50°C for 30min; use an ultrasonic pulverizer to perform ultrasonic treatment in an ice bath, Ultrasonic conditions are: ultrasonic intensity 40%, ultrasonic 3s, interval 5s, total action 20min, so that the stem cell factor is fully ...

specific Embodiment 3

[0043] The same part of this embodiment and specific embodiment 1 will not be repeated, the difference is:

[0044] Weigh 0.4g of soybean lecithin, 0.05g of cholesterol and 0.1g of Tween 80 and dissolve them in 10mL of absolute ethanol, and ultrasonically dissolve them fully; The mixture was introduced into 10 mL of 10% ammonium sulfate solution, and then placed in a round bottom flask; after the solution was evaporated under reduced pressure at 60°C to remove the organic solvent, it was placed in DPBS with pH 7.0 and dialyzed for 24 hours to form liposomes.

[0045] Put the dialyzed liposomes in a water bath shaker at 60°C for oscillation, then slowly add 20ml of the prepared stem cell extract, after the addition is completed, continue to shake at 60°C for 30min; use an ultrasonic pulverizer to perform ultrasonic treatment in an ice bath, Ultrasonic conditions are: ultrasonic intensity 40%, ultrasonic 3s, interval 5s, total action 20min, so that the stem cell factor is fully ...

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Abstract

The invention discloses a preparation method of stem cell factor liposome for repairing damaged tissues. The preparation method comprises the following steps: step 1, separating and culturing canine adipose-derived mesenchymal stem cells; 2, digesting the cultured canine adipose tissue-derived stromal cells, crushing, centrifuging, filtering and carrying out ultrafiltration to obtain a stem cell extracting solution; step 3, mixing the lipidosome prepared in advance with the stem cell extracting solution; carrying out ultrasonic treatment to obtain a stem cell factor liposome suspension, addingsodium propionate, uniformly mixing, and filtering to obtain stem cell factor liposome solution. The method has the advantages that the absorption rate of the stem cell factors can be increased, theactivity of the stem cell factors can be protected, the loss of effective components can be reduced, and the utilization rate of the effective components can be effectively increased. Meanwhile, the invention also discloses the stem cell factor liposome for repairing damaged tissues, which is prepared by the method, and stem cell factors in the preparation are pure natural components, so that thepreparation is safe to apply, free from toxic and side effects, free from drug residues, high in activity, small in clinical dosage and quick to take effect.

Description

technical field [0001] The invention belongs to the technical field of stem cells, in particular to a stem cell factor liposome for repairing damaged tissues and a preparation method thereof. Background technique [0002] Adipose-derived mesenchymal stem cells are a cell population with self-renewal and multi-directional differentiation potential in adipose tissue. Due to their convenient acquisition, multi-directional differentiation ability and strong in vitro proliferation ability, they have played an increasingly important role in wound healing and tissue repair in recent years. attention of researchers. Adipose-derived mesenchymal stem cells can actively migrate to the site of tissue damage and participate in immune regulation and tissue damage repair. The paracrine action of adipose-derived mesenchymal stem cells can produce a large amount of soluble stem cell factors to promote the repair of tissue damage. Mesenchymal stem cells participate in the main mechanism of t...

Claims

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Application Information

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IPC IPC(8): A61K35/28A61K9/127A61K47/26A61P17/02
CPCA61K9/127A61K35/28A61K47/26A61P17/02
Inventor 邢杰乔彦良苗昱韩爽李朝阳马广斌邹海涛李广森
Owner 青岛信迪细胞生物科技开发有限公司
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