Control of pH in human long-term bone marrow cultures with low-glucose medium containing zwitterion buffer lengthens the period of haemopoietic activity.
Primary long-term bone marrow cultures grown in 40 mM HEPES-buffered McCoy's 5A medium produced granulocyte-macrophage colony-forming units (CFU-GM) for a median of 9 weeks compared with 7 weeks with CO2/bicarbonate-buffered cultures. Reducing the medium glucose concentration (from 12.5 to 2.75 mM) extended the culture longevity to 17 weeks. The median period of erythroid colony detection increased from 6 to 8 weeks. Secondary cultures (5 x 106 cord blood mononuclear cells seeded on irradiated stroma) showed statistically similar myeloid and erythroid longevity to primary cultures. Improved control of medium pH significantly improved the capacity of long-term stromal layers to maintain stem cells in vitro.
Tennant
G B
GB
Department of Haematology, University of Wales College of Medicine, Heath Park, Cardiff, UK. tennant@cardiff.ac.uk
Truran
L N
LN
Bailey-Wood
R
R
Burnett
A K
AK
D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects; D002021:Buffers; D002454:Cell Differentiation; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D003470:Culture Media; D005947:Glucose / Q000494:pharmacology; D006410:Hematopoiesis / Q000187:drug effects*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D013997:Time Factors
D002021:Buffers; D003470:Culture Media; D005947:Glucose