Abstract
Clinical hematopoietic stem/progenitor cell (HSPC) transplantation outcomes are strongly correlated with the number of cells infused. Hence, to generate sufficient HSPCs for transplantation, the best culture parameters for expansion are critical. It is generally assumed that the defined oxygen (O2 ) set for the incubator reflects the pericellular O2 to which cells are being exposed. Studies have shown that low O2 tension maintains an undifferentiated state, but the expansion rate may be constrained because of limited diffusion in a static culture system. A combination of low ambient O2 and dynamic culture conditions has been developed to increase the reconstituting capacity of human HSPCs. In this unit, the protocols for serum-free expansion of HSPCs at 5% and 20% O2 in static and dynamic nutrient flow mode are described. Finally, the impact of O2 tension on HSPC expansion in vitro by flow cytometry and colony forming assays and in vivo through engraftment using a murine model is assessed.
| Original language | English |
|---|---|
| Pages (from-to) | 2A.11.1-2A.11.13 |
| Number of pages | 13 |
| Journal | Current Protocols in Stem Cell Biology |
| Volume | 44 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 28 Feb 2018 |
Keywords
- bone marrow repopulation assay
- dynamic culture system
- hematopoietic stem/progenitor cell expansion
- pericellular oxygen
- umbilical cord blood
Cite this
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Controlling the Effective Oxygen Tension Experienced by Cells Using a Dynamic Culture Technique for Hematopoietic Ex Vivo Expansion. / Tiwari, Abhilasha; Wong, Cynthia; Nekkanti, Lakshmi Prasanna; Deane, James Antony; McDonald, Courtney; Li, Jingang; Pham, Yen; Sutherland, Amy Elizabeth; Jenkin, Graham; Kirkland, Mark.
In: Current Protocols in Stem Cell Biology, Vol. 44, No. 1, 28.02.2018, p. 2A.11.1-2A.11.13.Research output: Contribution to journal › Article › Other › peer-review
TY - JOUR
T1 - Controlling the Effective Oxygen Tension Experienced by Cells Using a Dynamic Culture Technique for Hematopoietic Ex Vivo Expansion
AU - Tiwari, Abhilasha
AU - Wong, Cynthia
AU - Nekkanti, Lakshmi Prasanna
AU - Deane, James Antony
AU - McDonald, Courtney
AU - Li, Jingang
AU - Pham, Yen
AU - Sutherland, Amy Elizabeth
AU - Jenkin, Graham
AU - Kirkland, Mark
PY - 2018/2/28
Y1 - 2018/2/28
N2 - Clinical hematopoietic stem/progenitor cell (HSPC) transplantation outcomes are strongly correlated with the number of cells infused. Hence, to generate sufficient HSPCs for transplantation, the best culture parameters for expansion are critical. It is generally assumed that the defined oxygen (O2 ) set for the incubator reflects the pericellular O2 to which cells are being exposed. Studies have shown that low O2 tension maintains an undifferentiated state, but the expansion rate may be constrained because of limited diffusion in a static culture system. A combination of low ambient O2 and dynamic culture conditions has been developed to increase the reconstituting capacity of human HSPCs. In this unit, the protocols for serum-free expansion of HSPCs at 5% and 20% O2 in static and dynamic nutrient flow mode are described. Finally, the impact of O2 tension on HSPC expansion in vitro by flow cytometry and colony forming assays and in vivo through engraftment using a murine model is assessed.
AB - Clinical hematopoietic stem/progenitor cell (HSPC) transplantation outcomes are strongly correlated with the number of cells infused. Hence, to generate sufficient HSPCs for transplantation, the best culture parameters for expansion are critical. It is generally assumed that the defined oxygen (O2 ) set for the incubator reflects the pericellular O2 to which cells are being exposed. Studies have shown that low O2 tension maintains an undifferentiated state, but the expansion rate may be constrained because of limited diffusion in a static culture system. A combination of low ambient O2 and dynamic culture conditions has been developed to increase the reconstituting capacity of human HSPCs. In this unit, the protocols for serum-free expansion of HSPCs at 5% and 20% O2 in static and dynamic nutrient flow mode are described. Finally, the impact of O2 tension on HSPC expansion in vitro by flow cytometry and colony forming assays and in vivo through engraftment using a murine model is assessed.
KW - bone marrow repopulation assay
KW - dynamic culture system
KW - hematopoietic stem/progenitor cell expansion
KW - pericellular oxygen
KW - umbilical cord blood
UR - http://www.scopus.com/inward/record.url?scp=85059243310&partnerID=8YFLogxK
U2 - 10.1002/cpsc.42
DO - 10.1002/cpsc.42
M3 - Article
VL - 44
SP - 2A.11.1-2A.11.13
JO - Current Protocols in Stem Cell Biology
JF - Current Protocols in Stem Cell Biology
SN - 1941-7322
IS - 1
ER -