Biotechnology and Bioinformatics (1+1)

3.1.7. Flow chart for primary cell culture from fin fish

Fish

Swab with 70% alcohol or betadine to sterilize the external surfaces

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Remove caudal fin, gills and scales aseptically

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Cut the tissues in to rate fine pieces aseptically

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Wash the tissues with phosphate bufferd saline (PBS) 2-3 times

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Place the washed tissue in a sterile China dish or tissue culture flask and add 1-5 ml of Leibowitz’s L-15 medium, until the tissue is just submerged.

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Incubate at 28^oC – 29^oC for 24-72 hrs. The cells from the explant migrate into the surrounding medium and form a confluent monolayer

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A confluent monolayer may be formed in 3-4 days