STIRRED CULTURE OF PERIPHERAL AND CORD-BLOOD HEMATOPOIETIC-CELLS OFFERS ADVANTAGES OVER TRADITIONAL STATIC SYSTEMS FOR CLINICALLY RELEVANT APPLICATIONS

The ability to culture hematopoietic cells in readily characterizable and scalable stirred systems, combined with the capability to utilize serum-free medium, will aid the development of clinically attractive b ioreactor systems for transplantation therapies. We thus examined the proliferation and differentiation characteristics of peripheral blood (PB) mononuclear cells (MNC), cord blood (CB) MNC, and PB CD34(+) cell s in spinner flasks and (control) T-flask cultures in both serum-conta ining and serum-free media. Hematopoietic cultures initiated from all sources examined (PB MNC, CB MNC, and PB CD34(+) cells) grew well in s pinner vessels with either serum-containing or serum-free medium. Cult ure proliferation in spinner flasks was dependent on both agitator des ign and RPM as well as on the establishment of critical inoculum densi ties (ID) in both serum-containing (2 x 10(5) MNC/mL) and serum-free ( 3 x 10(5) MNC/mL) media. Spinner flask culture of PB WING in serum-con taining medium provided superior expansion of total cells and colony-f orming cells (CFC) at high ID (1.2 x 10(6) cells/mL) as compared to T- flask controls. Serum-free spinner culture was comparable, if not supe rior, to that observed in serum-containing medium. This is the first r eport of stirred culture of PB or CB MNC, as well as the first report of stirred CD34(+) cell culture. Additionally, this is the first accou nt of serum-free stirred culture of hematopoietic cells from any sourc e. (C) 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59: 534-543, 199 8.