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Separate fresh tissue of interest.
Treat tissue with enzyme(s) (e.g. trypsin, collagenase, protease) and/or mechanically to isolate cells.
Wash, count, and seed cells.
Examine cells under a brightfield microscope to assess their growth state, attachment to culture vessels/flasks, and to check for any signs of infection.
Monitor cells for the following days until they reach confluence.
Verify isolated cell types by their morphology and expressed biomarkers.
Passage cells to propagate the cell line.
Make master and working cell banks.
Immortalize cells if necessary.
Plan and execute experiments.
Keep monitoring cell state and possible infections using a brightfield microscope.