Method for preparing calcium alginate-fat acellular matrix microcarrier by applying micro-fluidic chip

Abstract

The invention provides a method for preparing a calcium alginate-fat acellular matrix microcarrier by applying a micro-fluidic chip. The method specifically comprises the following steps: subjecting human adipose tissue to repeated freeze thawing, performing emulsifying to obtain nano fat, performing centrifuging to remove upper-layer oil and lower-layer water, subjecting the middle layer to decellularization with a PBS buffer solution, a trypsin-EDTA digestive solution, isopropanol and a mixed enzyme solution, performing soaking, extracting and centrifuging with an alpha-amylase aqueous solution and acetic acid to obtain supernatant, mixing the supernatant with a sodium alginate solution with the same volume, taking the obtained mixture as a dispersed phase, introducing the mixture into amicro-fluidic chip, introducing n-decanol containing span-80 as a continuous phase into micro-fluidic chip to obtain acellular matrix-sodium alginate droplets coated by the n-decanol, finally introducing an n-decanol solution containing CaCl2 for crosslinking overnight, and washing off n-decanol to obtain the calcium alginate-fat acellular matrix microcarrier. The method can produce the acellularmatrix microcarrier with uniform components, and the obtained microcarrier is more beneficial to cell growth and can be used for culturing adipose-derived stem cells.

Description

technical field [0001] The invention belongs to tissue engineering biological scaffold microcarrier, in particular to a method for constructing a calcium alginate-fat acellular matrix microcarrier using a microfluidic chip, which can increase the adhesion of fat stem cells, and is a novel microcarrier. Background technique [0002] Soft tissue defect is a common clinical problem. Existing methods such as flap transplantation, fat filling, and hyaluronic acid filling have disadvantages such as trauma, easy absorption of fillers, and inflammatory reactions. The method of tissue engineering combines seed cells with bio-scaffold materials for filling, which can largely avoid these problems. However, currently commonly used bio-scaffold materials such as chitosan, fibrin, and hyaluronic acid do not have the ability to promote stem cell adhesion and proliferation. , The ability to differentiate. [0003] Acellular adipose matrix is ​​the remaining part of adipose tissue after ce...

AI Technical Summary

Problems solved by technology

However, the fat acellular matrix has a flocculent structure, which is difficult to prepare as a carrier

Microfluidic chips can manipulate liquids on a microscopic scale, and liquid droplets with uniform size and composition can be prepared by using liquid shear force; calcium alginate is a commonly used degradable material in the preparation of microcarriers by microfluidic technology, but its cell affinity Suboptimal, difficult to use for stem cell culture

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