I am isolating the human thymocytes from human thymus by mechnical dispersion by using both RPMI and PBS, no doubt , I am observing huge number of cells under microscope , but after 5 to 10 minutes, all the cells are getting lysed quickly. Anybody can suggest me what is the best method to isolate human thymocytes from thymus and how to sort it out this lysis problem and how to maintin health thymocytes after isolation and what is the other best method for isolation of thymocytes from thymus?
Many thymocytes are always on the verge of death anyway, and you don't mention the age of the individual(s) from which you acquired your thymocytes unfortunately, so it is likely that your starting material of thymocytes is unhealthy to begin with and the trauma of mechanical isolation and general manipulation renders them viably irretrievable.
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