BACKGROUND: The yield of white blood cells (WBCs) extracted from whole-blood leukoreduction filters can be affected by the storage conditions and delay before filtration. Platelets (PLTs) collected with apheresis instruments (Trima Accel, Gambro BCT) are leukoreduced during the procedure on a fluidized particle bed in a leukoreduction chamber (LRS chamber). In this report, the residual cell content of these LRS chambers was characterized to determine whether it would be a valuable source of viable human blood cells. STUDY DESIGN AND METHODS: The content of LRS chambers was eluted by gravity, and peripheral blood mononuclear cells (PBMNCs) were purified on a Ficoll-Paque gradient. Analyses were performed before and after freezing. Proportions of CD3+, CD14+, CD16+, CD19+, CD34+, and CD45+ cells were determined by flow cytometry. The frequency of T cells expressing CD4, CD8, and CD27 and of B cells expressing immunoglobulin G (IgG), IgM, and CD27 was also determined. RESULTS: LRS chambers held approximately 10(9) CD45+ cells representing the normal proportions of CD3+, CD14+, CD16+, and CD19+ cell populations of PBMNCs. A small fraction of these CD45+ cells were CD34+CD38+ cells (0.3 +/- 0.2%). The viability of these cells, measured before and after freezing, was more than 95 percent. CONCLUSION: The residual cell content of Trima Accel LRS chambers recovered after PLT collection is a good source of viable monocytes and lymphocytes. These PBMNCs, containing CD3+, CD14+, CD16+, CD19+, and CD34+ cells can be frozen to prepare cell banks, which opens new avenues for utilization in several physiologic studies or even in cellular therapy applications.
BACKGROUND: The yield of white blood cells (WBCs) extracted from whole-blood leukoreduction filters can be affected by the storage conditions and delay before filtration. Platelets (PLTs) collected with apheresis instruments (Trima Accel, Gambro BCT) are leukoreduced during the procedure on a fluidized particle bed in a leukoreduction chamber (LRS chamber). In this report, the residual cell content of these LRS chambers was characterized to determine whether it would be a valuable source of viable human blood cells. STUDY DESIGN AND METHODS: The content of LRS chambers was eluted by gravity, and peripheral blood mononuclear cells (PBMNCs) were purified on a Ficoll-Paque gradient. Analyses were performed before and after freezing. Proportions of CD3+, CD14+, CD16+, CD19+, CD34+, and CD45+ cells were determined by flow cytometry. The frequency of T cells expressing CD4, CD8, and CD27 and of B cells expressing immunoglobulin G (IgG), IgM, and CD27 was also determined. RESULTS: LRS chambers held approximately 10(9) CD45+ cells representing the normal proportions of CD3+, CD14+, CD16+, and CD19+ cell populations of PBMNCs. A small fraction of these CD45+ cells were CD34+CD38+ cells (0.3 +/- 0.2%). The viability of these cells, measured before and after freezing, was more than 95 percent. CONCLUSION: The residual cell content of Trima Accel LRS chambers recovered after PLT collection is a good source of viable monocytes and lymphocytes. These PBMNCs, containing CD3+, CD14+, CD16+, CD19+, and CD34+ cells can be frozen to prepare cell banks, which opens new avenues for utilization in several physiologic studies or even in cellular therapy applications.
Authors: John M Gansner; Mahboubeh Rahmani; A Helena Jonsson; Brooke M Fortin; Idayat Brimah; Martha Ellis; Robin Smeland-Wagman; Zhihan J Li; Jason M Schenkel; Michael B Brenner; Revital Yefidoff-Freedman; Steven R Sloan; Nancy Berliner; Nicolas C Issa; Lindsey R Baden; Dan L Longo; Duane R Wesemann; Donna Neuberg; Deepak A Rao; Richard M Kaufman Journal: Blood Date: 2018-11-14 Impact factor: 22.113