Low-oxygen and high-carbon-dioxide atmosphere improves the conservation of hematopoietic progenitors in hypothermia.
BACKGROUND
During short-term storage of hematopoietic cells (HCs) at 4°C a substantial decline in number and in functional capacity of progenitors occurs after 3 days. We hypothesized that physiologic O2 and CO2 concentrations of hematopoietic tissue microenvironment (approx. 3% O2 and approx. 6% CO2) could improve cell viability and functionality during storage at 4°C.


METHODS
Mobilized peripheral blood (PB) CD34+ cells from multiple myeloma or non-Hodgkin's lymphoma patients were stored in flasks containing air (approx. 20% O2 and approx. 0.05% CO2) or 3% O2/6% CO2 atmosphere, for 3, 5, and 7 days at 4°C. The total number of cells, the number of cells in G0 or G1 phase of cell cycle, and the apoptosis rate were determined. The functional capacity of stored cells was assessed by the capacity of progenitors to form colonies in methylcellulose (colony-forming cells [CFCs]) and of stem cells to repopulate the bone marrow (BM) of immunodeficient mice (SCI D-repopulating cell [SRC] assay).


RESULTS
The total number of viable cells and cells in G1 phase as well as the number of total CFCs were significantly higher at 3% O2/6% CO2 than in air at all time points. Cells in G0 phase and SRC were equally preserved in both conditions.


CONCLUSIONS
Atmosphere with low O2 and high CO2 concentration (3% O2/6% CO2) in hypothermia (+4°C) during 7 days of storage prevents cell damage and preserves a high number of functional HSCs and progenitors mobilized in PB by granulocyte-colony-stimulating factor.
Jeanne
Michel
M
Aquitaine-Limousin Branch of French Blood Institute, Bordeaux, France.
Kovacevic-Filipovic
Milica
M
Szyporta
Milène
M
Vlaski
Marija
M
Hermitte
Francis
F
Lafarge
Xavier
X
Duchez
Pascale
P
Boiron
Jean-Michel
JM
Praloran
Vincent
V
Ivanovic
Zoran
Z
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002245:Carbon Dioxide / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D005260:Female; D016193:G1 Phase / Q000187:drug effects; D019650:Hematopoietic Stem Cell Mobilization; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D008228:Lymphoma, Non-Hodgkin / Q000378:metabolism; D008297:Male; D051379:Mice; D016513:Mice, SCID; D009101:Multiple Myeloma / Q000378:metabolism; D010100:Oxygen / Q000494:pharmacology*; D036102:Peripheral Blood Stem Cell Transplantation; D012034:Refrigeration; D016192:Resting Phase, Cell Cycle / Q000187:drug effects; D014183:Transplantation, Heterologous
D002245:Carbon Dioxide; D010100:Oxygen