Dear all list members I posted a question about the lymphocyte subset problem after CD34 selection. Now, I'll tell you more about it. 1. Dr. Timothy Farley asked me about the absolute number and the proportion of CD34 positive cells. -> First day PBPC collection bag : 1233/uL (0.92%) Post CD34 selection : 1172/uL (35.0%) Second day PBPC collection bag : 962/uL (0.45%) Post CD34 selection : 550/uL (45.0%) 2. Dr. Dennis told to me that anti-CD19 antibody might stick to CD34 positive cells and their fluorescence also might be dim. He said in his experience that bright CD19 (true) cells are about 1%. -> Our results showed that CD19 fluorescence was as bright as other fluorescences such as CD3 and CD19 positive populations were identified without difficulty. 3. Dr. Donnenberg said that CD3+/CD19+ cells could be T cell/ B cell conjugates. -> I can't imagine all CD3+ and CD3- cells might conjugate with CD19+ B c ells. 4. I tested the possibility that some procedure could make all cells change their unknown surface antigen and then they got the power to bind anti-CD19 antibody. -> I tried to simulate all procedure such as wash, incubation, and so on for the selection of CD34 positive cells without anti-CD34 monoclonal antibody and passing through the column which captures the CD34 positive cells. Unfortunately, the lymphocyte subset was normal. Now, I have no way to solve the problem other than waiting another PBPC and CD34 selection. Is there anyone who can give me a hint? Thanks a lot. ------------------------------ Chang-Seok Ki, MD Dept. of Clinical Pathology Samsung Medical Center Seoul, Korea Voice (82)-2-3410-2708 Fax (82)-2-3410-2719 E-mail kcdol@samsung.co.kr -----------------------------
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