Serum-free culture conditions for cells capable of producing long-term survival in lethally irradiated mice

Stem Cells
R L BrownM Patchen

Abstract

The goal of ex vivo culture is to expand and/or differentiate cells in culture such that they retain their functional characteristics when reinfused into a patient. The studies presented here analyzed the use of culture conditions devoid of serum to expand murine hematopoietic stem cells. Bone marrow cells from male B6D2F1/J mice were cultured for up to 28 days in serum-free medium in the absence or presence of stem cell factor (SCF), GM-CSF or a combination of the two factors. Cells cultured for up to 21 days were assessed for granulocyte-macrophage colony-forming cells (GM-CFC), spleen colony-forming units, and cells responsible for short-term and long-term hematopoietic repopulation in lethally irradiated mice. Compared to initial seeding levels, the presence of SCF and GM-CSF increased total cell numbers 90-fold and GM-CFC numbers 42-fold over a 21-28 day culture period. Although spleen colony-forming unit cells did not increase, they were maintained at initial seeding levels over a 21-day period in the presence of SCF and GM-CSF. In lethally irradiated mice, survival enhancement and hematologic reconstitution were optimum with cells cultured for only seven days: survival at six months was 100% with cells cultured in SCF pl...Continue Reading

References

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Citations

Nov 17, 2009·In Vitro Cellular & Developmental Biology. Animal·Jinhyung ParkPeter C W Kim
Sep 12, 2000·Journal of Hematotherapy & Stem Cell Research·J H WonH S Park
Jul 6, 2000·The Orthopedic Clinics of North America·J S GreenbergerR K Houck
May 18, 2016·Pharmaceutical Research·Timothy HammondHolly Birdsall

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