We are attempting to isolate lymphocytes for Rhesus lymph nodes, but have a clumping problem. Currently, we grind the nodes in a sterile tissue grinder, filter the cell suspension to remove garbage, and then wash. Most of the cells clump after the first centrifugation. We have tried several washing solutions (PBS with and without albumin, with and without EDTA, RPMI with and without albumin etc) and multiple RCF and the clumping always occurs. The procedure requires the washing steps so we are left in a bind. Any help or recommendations will be greatly appreciated. Paul Kuon MT(ASCP) Research Associate University of Louisiana Lafayette NIRC
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