Preparation method of extracellular matrix-imitated printable composite biological ink

Abstract

The invention discloses a preparation method of extracellular matrix-imitated printable composite biological ink, which comprises the following steps of: (A) dissolving GelMA powder and a photoinitiator in PBS, and uniformly stirring at 40-50 DEG C to prepare a GelMA solution with the final concentration of 5-15%; (B) pretreating fresh adipose tissues, mashing the pretreated adipose tissues, respectively treating the mashed adipose tissues by 0.25% trypsin-0.1% EDTA (Ethylene Diamine Tetraacetic Acid) and 3% Triton-X100 for 1 hour, and respectively treating the treated adipose tissues by 4% sodium deoxycholate and 100% isopropanol; (C) dissolving the acellular extracellular matrix obtained in the step (B) into 0.01 M HCL and 1mg / mL pepsin solution, stirring at 4-10 DEG C, and centrifugingat a low speed for 10min to obtain a deadipose extracellular matrix solution with the final concentration of 5-15mg / mL; and (D) preheating the GelMA prepared in the step (A) at a proper temperature, slowly adding the solution in the step (C) at a vortex oscillation speed of 1500-2500rpm according to a volume ratio of the GelMA to the acellular extracellular matrix of (1-4): 1, adjusting the pH value, and performing sterilizing. The composite biological ink prepared by the method has excellent printability, biocompatibility and biological activity.

Description

technical field [0001] The invention relates to a preparation method of imitating extracellular matrix and printable composite biological ink. Background technique [0002] The traditional two-dimensional cell culture is quite different from the real tumor microenvironment due to the inability to imitate the interaction between cells in the three-dimensional environment of the tumor and the lack of corresponding extracellular matrix (ECM); and the large racial differences between the animal model and the human body are difficult to achieve. It reflects the real tumor environment of the human body, and there are ethical issues. Therefore, there are deviations in the clinical understanding of tumor pathogenesis, treatment and metastasis. The bionic three-dimensional tumor model containing extracellular matrix and a variety of active cells can provide a reliable platform for intuitive research on tumor development, metastasis, anti-tumor drug development and screening in vitro ...

AI Technical Summary

Problems solved by technology

In terms of printability, existing bioinks still have disadvantages such as poor injectability, slow curing speed, low mechanical strength, and uncontrollable degradation performance; in terms of biological activity, existing bioinks have low bioactivity and lack the ability to promote cell proliferation and differentiation ECM-related biochemical signals and specific functional expression, difficult to generalize the culture conditions of real cells

3D bioprinting to construct in vitro biomimetic 3D tumor models remains challenging due to the lack of bioinks with both well-printed and cell-mimicking components

The acellular extracellular matrix material maintains the microenvironment of cell growth, including collagen, protein, polysaccharide and physical and chemical factors, but its poor formability is difficult to be printed, so it needs to be compounded with materials with better curing properties, such as excellent biophase Capacitance and fast photocuring capability of gelatin methacrylic anhydride (GelMA)