Adipose tissue-derived composite cell group and preparation method and application thereof

Abstract

The invention discloses an adipose tissue-derived composite cell group and a preparation method and an application thereof. The preparation method of the adipose tissue-derived composite cell group includes the following steps: (1) washing adipose tissue; (2) separating the adipose tissue, wherein the washed adipose tissue sequentially passes through three blade-type nano adipose transplantation emulsifiers with different pore diameters of 1.5 mm-2.0 mm, 0.8 mm-1.0 mm and 0.5 mm-0.7 mm, removing an oil drop layer, a mature adipose cell layer and a physiological saline layer after centrifugation, and collecting the composite cell group layer at the bottom; filtering the composite cell group layer with a cell sieve to remove cell fragments and base substance components in the composite cellgroup layer, centrifuging, and discarding supernatant to obtain the adipose tissue-derived composite cell group. The adipose tissue is crushed under the cutting action, the acting force on the adiposetissue is reduced, and damage to target products such as mesenchymal stem cells and the like is avoided, so that the cell structure is protected, and the treatment effect of the products is improved.

Description

technical field [0001] The invention belongs to the field of stem cell therapy, and in particular relates to a compound cell group derived from adipose tissue and its preparation method and application. Background technique [0002] In the prior art, adipose tissue-derived composite cell populations are prepared by enzymatic digestion, but there are many deficiencies in the enzymatic digestion method, which limits the clinical application of multicellular components derived from adipose tissue, as follows: [0003] 1. Collagenase (bovine origin) and collagenase combined with trypsin (pig origin) are commonly used for enzymatic digestion to digest collagen and protein between fat cells to obtain the cell components of the adipose tissue-derived compound cell group, and use bovine Serum albumin (BSA) terminates the reaction of trypsin and uses animal-derived digestive enzymes, which is complicated to operate, high in cost, and poses a biological safety hazard. [0004] 2. In ...

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Problems solved by technology

[0002] The prior art uses the enzyme digestion method to prepare composite cell populations derived from adipose tissue, but there are many shortcomings in the enzyme digestion method, which limits the clinical application of multicellular components derived from adipose tissue, as follows:

[0003] 1. Collagenase (bovine origin) and collagenase combined with trypsin (pig origin) are commonly used for enzymatic digestion to digest collagen and protein between fat cells to obtain the cell components of the compound cell group derived from adipose tissue, and use bovine Serum albumin (BSA) terminates the reaction of trypsin and uses animal-derived digestive enzymes, which is complicated to operate, high in cost, and poses a biological safety hazard

[0004] 2. Enzyme digestion method needs to add human serum albumin during separation in order to ensure cell viability, using drugs from scarce human resources, and the cost is high

[0005] 3. A large number of red clumps can be seen in the cells separated by enzymatic digestion, and a large number of round cells with uniform shape can be seen under the microscope. The current method to remove red blood cell contamination is to use red blood cell lysate and use the principle of hypotonicity to lyse red blood cells, or use lymphocytes The separation liquid is passed through the density gradient centrifugation method, the reagents used in the two methods are not medical products, the red blood cell pollution is serious, and the cost is high

Similarly, ultrasonic cavitation destroys cells indiscriminately, damages target products such as mesenchymal stem cells, destroys cell structure, and affects the therapeutic effect of products

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