Method for efficiently separating human synovium primary cells
A technology of primary cells and synovium, applied in the field of biotechnology and cell culture, can solve the problems of inability to effectively digest and separate fibrous tissue, long growth cycle of primary cells, and difficulty in dissociation of synoviocytes, etc., to simplify the primary culture method , shorten the time of sticking to the wall, and improve the efficiency of scientific research
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Embodiment 1
[0042] Take 6.7g of human fresh synovial tissue (observe loose connective tissue, light red, smooth and shiny surface, thin and soft, with fluffy brush shape on the edge) and put it in a sterile petri dish with a diameter of 10cm, and mix it with Hank's balanced salt The solution was repeatedly washed 4-5 times until the residual blood was rinsed away. Transfer the tissue to a sterile culture bottle with a diameter of 3.5 cm, and use sterile scissors to cut the synovial tissue into pieces for 40-60 minutes, until the pieces are muddy, non-granular, and the size is not greater than 1 mm 3 ; Use a 3ml sterile Pasteur tube to transfer to the EP tube, then wash the petri dish with Hank's balanced salt mixture, then transfer it to the EP tube, and place it in a centrifuge for centrifugation at 1500rpm for 5min. Remove the supernatant from the precipitate after centrifugation, add 3ml of DMEM culture solution containing 1% penicillin and streptomycin mixed solution, and add 300μl of...
Embodiment 2
[0046] Take 5.2g of human fresh aseptic synovial tissue (observe loose connective tissue, light red, smooth and shiny surface, thin and soft, with fluffy brush shape on the edge) and put it in a sterile petri dish with a diameter of 10cm, and use Hank's balance The salt mixture was washed repeatedly 45 times until the residual blood was rinsed away. Transfer the tissue to a sterile culture bottle with a diameter of 3.5 cm, cut the synovial tissue with sterile scissors for 40-60 min, it will be muddy, without grain feeling, and the volume size is not greater than 1 mm 3 ; Use a 3ml sterile Pasteur tube to transfer to the EP tube, then wash the petri dish with Hank's balanced salt mixture, then transfer it to the EP tube, and place it in a centrifuge for centrifugation at 1500rpm for 5min. After centrifugation, the supernatant was removed from the precipitate, and the precipitate was added to 3ml of DMEM culture solution containing 1% penicillin and streptomycin mixed solution, ...
Embodiment 3
[0050] Take 5.6g of human fresh aseptic synovial tissue (observe loose connective tissue, light red, smooth and shiny surface, thin and soft, with fluffy brush shape on the edge) and put it in a sterile petri dish with a diameter of 10cm, and use Hank's balance The salt mixture was washed repeatedly 4-5 times until the residual blood was rinsed away. Transfer the tissue to a sterile culture bottle with a diameter of 3.5 cm, cut the synovial tissue with sterile scissors for 40-60 minutes, and it will be muddy, without graininess, and the volume size is not greater than 1mm 3 ; Use a 3ml sterile Pasteur tube to transfer to the EP tube, then wash the petri dish with Hank's balanced salt mixture, then transfer it to the EP tube, and place it in a centrifuge for centrifugation at 1500rpm for 5min. The precipitate after centrifugation was removed from the supernatant, and the precipitate was added to 3ml of DMEM culture solution containing 1% penicillin and streptomycin mixed soluti...
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