Canoidea dermal fibroblast cell primary culture method

Abstract

The invention belongs to the technical field of cell culture and in particular relates to a canoidea dermal fibroblast cell primary culture method. The canoidea dermal fibroblast cell primary culturemethod comprises the following steps: (1) scissoring a thoracoabdominal skin tissue of an adult dog; (2) cleaning the skin tissue, removing subcutaneous fat and connective tissues and cutting the skintissue into pieces and transferring the skin tissue to a centrifugal tube; (3) adding 1 mL II collagenase solution into the centrifugal tube and scissoring the solution into paste, adding 5 mL II collagenase, uniformly mixing, putting the solution in a culture box till tissue slurry is digested fully; (4) taking out the centrifugal tube for centrifugalization, abandoning a supernate, adding a culture medium to re-suspend, filtering the re-suspended solution with a cell screen, carrying out centrifugalization again on the filtrate, abandoning the supernate, adding a cell culture solution to re-suspend, then transferring the solution to a hexagonal porous plate, and adding the culture solution; and (5) putting the solution in the culture box to stand to be cultured for 48 hours, and replacing a fresh culture solution every day till the canoidea dermal fibroblast primary cells are obtained. The method is low in operating difficulty, simple in culture flow and relatively high in culture efficiency.

Description

Technical field [0001] The invention belongs to the technical field of cell culture, and specifically relates to a method for primary culture of canine dermal fibroblasts. Background technique [0002] Fibroblasts are the most common cells in connective tissues. They are derived from the mesoderm. They are large in size, have clear outlines, and are protruding spindle-shaped or star-shaped structures. They are usually in a relatively dormant state and mainly synthesize extracellular matrix and collagen. A degree of cell degeneration, necrosis, tissue defect and bone trauma repair play a very important role. After trauma, fibroblasts will enter the active proliferation phase, and participate in the repair process by secreting various cytokines and their own differentiation. At present, there are many researches on human skin fibroblasts, and the culture technology is relatively mature. The Chinese invention patent with publication number CN106591223A discloses a method for separa...

AI Technical Summary

Problems solved by technology

There have been reports on the culture of canine skin fibroblasts at home and abroad, but these methods are somewhat complicated in actual operation, and some have a low success rate

The Chinese invention patent with the publication number CN105441377A discloses a primary separation method of animal skin fibroblasts. The method takes the damaged skin area, cleans it and shreds it, then pours it into PBS with 1% double antibody, centrifuges, and inoculates Culture, although this method successfully increases the number of cells and is more conducive to cell climbing out, but the activity of the isolated cells is low, which is not conducive to subsequent subculture