Cell Culture Device for Hematopoietic Progenitor Cells

This research will examine growth of normal human marrow in a compact cell culture"OXYCELL" bioreactor. This bioreactor is suited for experimental manipulation due to its small cellvolume and in-house manufacture. This research will examine the benefit of providing an adherentsubstrate in the bioreactor for stromal cell attachment and growth. Two distinct substrate choices,polysulfone ultrafiltration membrane and "OPTICELL" ceramic matrix, will be tested and compared.Hematopoietic lineages culture will be quantified to assess performance differences. The second partdeals with the seeking of CD34+ progenitor cells into a bioreactor in which an established stromal cellmicroenvironment is compartmentalized within a 0.2 uM microfiltration membrane. The progenitor cellswill share cell space medium with the adherent cells but will not be in direct cell-cell contact. Growthwill be compared to cells seeded directly into the microenvironment space and into a bioreactor withoutan established microenvironment. Exogenous growth factors will be tested for impact on the growthof progenitor cells seeded into the bioreactors. Cultured cells will be phenotyped for lineage commitmentand differentiative stage and assayed for uncommitted and committed progenitors. This work is todevelop a bioreactor suited for bone marrow culture applications. The bioreactor is to be incorporatedinto a cultureware to be supported by an automated cell culture instrument (currently underdevelopment) designed specifically for clinical use in immunotherapy.