Ѳǰܱ™ - Long-Term Culture-Initiating Cell (LTC-IC) Assays
Ѳǰܱ™ Media for LTC-IC Assays
Long-Term Culture of Human and Mouse Hematopoietic Progenitor Cells
Ѳǰܱ™ media have been developed for the long-term culture of hematopoietic progenitor cells in association with adherent stromal cells. An important application of Ѳǰܱ™ is the quantitation of primitive hematopoietic progenitor cells in the
long-term culture-initiating cell (LTC-IC) assay
. In this assay hematopoietic cells are cultured in Ѳǰܱ™ medium with supportive human or mouse stromal cell layers and tested for their ability to generate myeloid clonogenic progenitors for several weeks as assayed by replating the cells in a
using
medium.
Why Use Ѳǰܱ™?
- Establish stromal cell layers and culture hematopoietic progenitors in “Dexter-type” long-term bone marrow cultures.1,2
- Perform LTC-IC assays in Ѳǰܱ™ media.3
- Study regulators of myelopoiesis in long-term cultures.3
Protocol

Figure 1. Procedure for Limiting-Dilution Long Term Culture-Initiating Cell (LTC-IC) Assay

Figure 2. Procedure for Bulk Culture LTC-IC Assay
Key Applications
Human Hematopoietic Cell Culture
Nemeth M et al. (2009) Stem Cells 27(5): 1109-1119
Tipping A et al. (2009) Blood
Peterson M et al. (2008) Immunity 29(4): 578-588
Carlo-Stella C et al. (2007) Stem Cells 25(1): 252-261
Pastos K et al. (2006) Blood 108(10): 3360-3362
Sloand E et al. (2006) Proc Natl Acad Sci 103(39): 14483-14488
Pirson L et al. (2006) Stem Cells 24(7): 1814-1821
Takeda A et al. (2006) Cancer Res 66(13): 6628-6637
Giebel B et al. (2006) Blood 107(5): 2146-2152
Srour E et al. (2005) Blood 105(8): 3109-3116
Selleri C et al. (2005) Blood 105(5): 2198-2205
Giebel B et al. (2004) Blood 104(8): 2332-2338
Kogler G et al. (2004) J of Exp Med 200(2): 123-135
Guan Y et al. (2003) Blood 101(8): 3142-3149
Bartolovic K et al. (2004) Blood 103(2): 523-529
Stier S et al. (2003) Blood 102(4): 1260-1266
Danet G et al. (2003) J of Clin Invest 112(1): 126-135
Dybedal I et al. (2003) Blood 102(1): 118-126
Punzel M et al. (2003) Exp Hematol 31(4): 339-347
Murdoch B et al. (2003) Proc Natl Acad Sci 100(6): 3422-3427
Kootstra N et al. (2003) Proc Natl Acad Sci U S A 100(3): 1298-1303
Bhatia M et al. (1998) Nat Med 4(9): 1038-1045
Dybedal I et al. (2003) Blood 102(1): 118-126
Tipping A et al. (2009) Blood
Peterson M et al. (2008) Immunity 29(4): 578-588
Carlo-Stella C et al. (2007) Stem Cells 25(1): 252-261
Pastos K et al. (2006) Blood 108(10): 3360-3362
Sloand E et al. (2006) Proc Natl Acad Sci 103(39): 14483-14488
Pirson L et al. (2006) Stem Cells 24(7): 1814-1821
Takeda A et al. (2006) Cancer Res 66(13): 6628-6637
Giebel B et al. (2006) Blood 107(5): 2146-2152
Srour E et al. (2005) Blood 105(8): 3109-3116
Selleri C et al. (2005) Blood 105(5): 2198-2205
Giebel B et al. (2004) Blood 104(8): 2332-2338
Kogler G et al. (2004) J of Exp Med 200(2): 123-135
Guan Y et al. (2003) Blood 101(8): 3142-3149
Bartolovic K et al. (2004) Blood 103(2): 523-529
Stier S et al. (2003) Blood 102(4): 1260-1266
Danet G et al. (2003) J of Clin Invest 112(1): 126-135
Dybedal I et al. (2003) Blood 102(1): 118-126
Punzel M et al. (2003) Exp Hematol 31(4): 339-347
Murdoch B et al. (2003) Proc Natl Acad Sci 100(6): 3422-3427
Kootstra N et al. (2003) Proc Natl Acad Sci U S A 100(3): 1298-1303
Bhatia M et al. (1998) Nat Med 4(9): 1038-1045
Dybedal I et al. (2003) Blood 102(1): 118-126
Mouse Hematopoietic Cell Culture
Zimmer S et al. (2011) Blood
Lymperi S et al. (2010) Blood 117(5): 1540-1549
Ohno Y et al. (2010) Proc Natl Acad Sci
Ohmori T et al. (2010) J Biol Chem 285(41): 31763-31773
Zhang Q et al. (2010) Blood
Sumitomo A et al. (2010) Mol Cell Biol
Pandit T et al. (2006) Stem cells 24(11): 2373-2381
Lee Y et al. (2006) Blood 108(7): 2416-2419
Carella C et al. (2006) Blood 107(3): 1124-1132
Carmen M et al. (2004) Blood 103(9): 3349-3354
Lacout C et al. (2003) Blood 102(4): 1282-1289
Truong B et al. (2003) Blood 101(3): 1141-1148
Jin H et al. (2002) J of Clin Invest 109(9): 1183-1191
Samper E et al. (2002) Blood 99(8): 2767-2775
Iversen P et al. (2002) Am J of Physiol, Regul, Integr Comp Physiol 282(1): R166-72
Jasinski M et al. (2001) Blood 98(7): 2248-2255
Lymperi S et al. (2010) Blood 117(5): 1540-1549
Ohno Y et al. (2010) Proc Natl Acad Sci
Ohmori T et al. (2010) J Biol Chem 285(41): 31763-31773
Zhang Q et al. (2010) Blood
Sumitomo A et al. (2010) Mol Cell Biol
Pandit T et al. (2006) Stem cells 24(11): 2373-2381
Lee Y et al. (2006) Blood 108(7): 2416-2419
Carella C et al. (2006) Blood 107(3): 1124-1132
Carmen M et al. (2004) Blood 103(9): 3349-3354
Lacout C et al. (2003) Blood 102(4): 1282-1289
Truong B et al. (2003) Blood 101(3): 1141-1148
Jin H et al. (2002) J of Clin Invest 109(9): 1183-1191
Samper E et al. (2002) Blood 99(8): 2767-2775
Iversen P et al. (2002) Am J of Physiol, Regul, Integr Comp Physiol 282(1): R166-72
Jasinski M et al. (2001) Blood 98(7): 2248-2255
Other Uses
Human NK Cell Culture
Keskin D et al. (2007) Proc Natl Acad Sci 104(9): 3378-3383
Chen X et al. (2006) Proc Natl Acad Sci 103(27): 10346-10351
Vieillard V et al. (2005) Proc Natl Acad Sci 102(31): 10981-10986
Baumann B et al. (2004) J Immunol 172(10): 6460-6467
Perez S et al. (2003) Blood 101(9): 3444-3450
Chen X et al. (2006) Proc Natl Acad Sci 103(27): 10346-10351
Vieillard V et al. (2005) Proc Natl Acad Sci 102(31): 10981-10986
Baumann B et al. (2004) J Immunol 172(10): 6460-6467
Perez S et al. (2003) Blood 101(9): 3444-3450
Human Mesenchymal Cell Culture
Chen X et al. (2006) Stem Cells 24(9): 2052-2059
Chagraoui J et al. (2003) Blood 101(8): 2973-2982
Baksh D et al. (2003) Exp Hematol 31(8): 723-732
Chagraoui J et al. (2003) Blood 101(8): 2973-2982
Baksh D et al. (2003) Exp Hematol 31(8): 723-732
References
- Dexter T et al. (1977) J Cell Physiol 91(3): 335–44
- Gartner S et al. (1980) Proc Natl Acad Sci 77(8): 4756–9
- Sutherland H et al. (1990) Proc Natl Acad Sci 87(9): 3584–8