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Effects of CD34+ cell selection and perfusion on ex vivo expansion of
peripheral blood mononuclear cells
CE Sandstrom, JG Bender, ET Papoutsakis and WM Miller
Department of Chemical Engineering, Northwestern University, Evanston, IL
60208-3120, USA.
Ex vivo expansion of peripheral blood mononuclear cells (MNCs), cultured
both directly and after selection for CD34+ cells, was compared in static
and continuously perfused cultures containing interleukin (IL)-3, IL-6,
granulocyte colony-stimulating factor (G- CSF), and stem cell factor (SCF).
Cultures inoculated with either MNCs or CD34+ cells produced cells that
were remarkably similar after 10 days of culture, as evidence by cell
morphology, expression of CD34, CD33, CD15, and CD11b, and the fractions of
cells giving rise to colony- forming units granulocyte-monocyte (CFU-GM)
and long-term culture- initiating cells (LTC-IC). Static and perfusion
cultures gave similar average total cells and CFU-GM expansions for both
MNC and CD34+ cell cultures. However, those samples that performed poorly
in static culture performed at near-normal levels in perfusion. In
addition, perfusion supported higher LTC-IC numbers for both MNC and CD34+
cell cultures. While total cell expansion was about ten times greater in
CD34+ cell cultures (approximately 100-fold), CFU-GM expansion
(approximately 20-fold) was similar for both MNC and CD34+ cell cultures.
The similar distribution of cell types produced in MNC and CD34+ cell
cultures allows direct comparison of total and colony- forming cell
production. After 15 days in perfusion, MNC cultures produced 1.5-, 2.6-,
and 2.1-fold more total cells, CFU-GM, and LTC-IC, respectively, than the
same sample selected and cultured as CD34+ cells. Even if the CD34+
selection process was 100% efficient, CFU-GM production would be 1.5-fold
greater for MNCs than for CD34+ cells.
Volume 86,
Issue 3,
pp. 958-970,
08/01/1995
Copyright © 1995 by The American Society of Hematology
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