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Post-thaw losses of progenitor and T cells from peripheral blood stem cells cryopreserved in a pentastarch solution do not increase for up to 2 hours after thawing

, : Post-thaw losses of progenitor and T cells from peripheral blood stem cells cryopreserved in a pentastarch solution do not increase for up to 2 hours after thawing. Blood 96(11 Part 1): 520a, November 16

Current practice guidelines dictate that thawing and infusion of cryopreserved PBSC be carried out as rapidly as possible. To support our institutional policies for thawing in the laboratory prior to transporting PBSC to clinical care units and to allow for unforeseen infusion delays, we studied the effect of extended room temperature (RT) storage on thawed PBSC. PBSC were collected on the Fenwal CS3000 from 12 G-CSF mobilized normal donors (12 L volume processed). Unseparated PBSC were cryopreserved in Cryocyte bags (Nexell) using a controlled rate freezer in 5% DMSO/6% pentastarch/4% HSA, placed in an overwrap bag, and stored in liquid nitrogen for 2 weeks. All 12 products were thawed rapidly in a 37degreeC water bath, diluted 1:4 with heparinized Plasmalyte A (Baxter), and held at RT for 120 min. Samples were obtained before cryopreservation and at 30 and 120 min after thawing, for cell counting, flow cytometric phenotyping and viability, and clonogenic assays. Values for content of total nucleated cells (TNC), CD34+ cells, CD3+ cells, and CFU-GM in the table represent mean (S.D.) of samples from 12 bags, each of which contained 1/4 of the initial product. Values for CD34+ and CD3+ cells represent viable (7AAD-neg) cells that express those markers. From pre-freeze to 30 min. post-thaw, expected losses occurred in % viability of the overall product (98% to 78%) and TNC (11.6 to 8.9 X 109). Loss of CD34+ cells was minimal (110 to 98 X 106; 89% recovery), but loss of CD3+ cells was substantial (4.0 to 1.6 X 109, 40% recovery). No significant further losses occurred between 30 min and 120 min for any of the assayed values. There were no significant changes in CFU-GM at any stage. Studies of T cell function are in progress to evaluate the post-thaw decline in assayed T cells. These data suggest that PBSC can be held at RT for up to 2 hours after thawing with no evident damage to progenitor and T cell populations beyond losses occurring after the initial freeze/thaw process.


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