Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow

DC Colter, R Class, CM DiGirolamo… - Proceedings of the …, 2000 - National Acad Sciences
DC Colter, R Class, CM DiGirolamo, DJ Prockop
Proceedings of the National Academy of Sciences, 2000National Acad Sciences
Cultures of plastic-adherent cells from bone marrow have attracted interest because of their
ability to support growth of hematopoietic stem cells, their multipotentiality for differentiation,
and their possible use for cell and gene therapy. Here we found that the cells grew most
rapidly when they were initially plated at low densities (1.5 or 3.0 cells/cm2) to generate
single-cell derived colonies. The cultures displayed a lag phase of about 5 days, a log
phase of rapid growth of about 5 days, and then a stationary phase. FACS analysis …
Cultures of plastic-adherent cells from bone marrow have attracted interest because of their ability to support growth of hematopoietic stem cells, their multipotentiality for differentiation, and their possible use for cell and gene therapy. Here we found that the cells grew most rapidly when they were initially plated at low densities (1.5 or 3.0 cells/cm2) to generate single-cell derived colonies. The cultures displayed a lag phase of about 5 days, a log phase of rapid growth of about 5 days, and then a stationary phase. FACS analysis demonstrated that stationary cultures contained a major population of large and moderately granular cells and a minor population of small and agranular cells here referred to as recycling stem cells or RS-1 cells. During the lag phase, the RS-1 cells gave rise to a new population of small and densely granular cells (RS-2 cells). During the late log phase, the RS-2 cells decreased in number and regenerated the pool of RS-1 cells found in stationary cultures. In repeated passages in which the cells were plated at low density, they were amplified about 109-fold in 6 wk. The cells retained their ability to generate single-cell derived colonies and therefore apparently retained their multipotentiality for differentiation.
National Acad Sciences