Source:http://linkedlifedata.com/resource/pubmed/id/10049066
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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1
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pubmed:dateCreated |
1999-2-25
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pubmed:abstractText |
The goal of this study was to evaluate if differences in culture conditions used in long-term culture assays affect enumeration of LTC-IC in freshly sorted or ex vivo expanded CD34+/HLA-DRdim/CD2-/CD7- (34+/Lin-) cells. The variables examined included different stromal feeders (murine bone marrow fibroblast cell line, M2-10B4 and murine fetal liver cell line, AFT024) and presence or absence of cytokines (MIP-1alpha + IL-3). The absolute LTC-IC frequency in 34+/Lin- cells measured in limiting dilution assays (LDA) on AFT024 (4.45 +/- 0.69%) was significantly higher than in M2-10B4 (1.45 +/- 0.20%) LDA. Addition of MIP-1alpha and IL-3 to AFT024 LDA increased the measured LTC-IC frequency to 6.8 +/- 0.9%. We also determined the fraction of LTC-IC that persisted after 34+/Lin cells were cultured for 5 weeks by replating progeny in the three LDA readout systems. The measured LTC-IC maintenance was significantly lower when M2-10B4 LDA (13.1 +/- 3.5%, P < 0.05) were used compared with AFT024 LDA (36.6 +/- 5.5%) or AFT024 LDA supplemented with MIP-1alpha and IL-3 (29.1 +/- 6.3%). Thus, the number of LTC-IC measured in freshly sorted 34+ cells depends on the stromal feeder used in LDA assays. Furthermore, and most important, assessment of LTC-IC expansion or maintenance may vary significantly depending on the type of stromal feeder used to enumerate LTC-IC.
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pubmed:grant | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Jan
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pubmed:issn |
0887-6924
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
13
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
92-7
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:10049066-Animals,
pubmed-meshheading:10049066-Antigens, CD,
pubmed-meshheading:10049066-Bone Marrow Cells,
pubmed-meshheading:10049066-Cell Culture Techniques,
pubmed-meshheading:10049066-Cells, Cultured,
pubmed-meshheading:10049066-Coculture Techniques,
pubmed-meshheading:10049066-HLA-DR Antigens,
pubmed-meshheading:10049066-Hematopoietic Stem Cells,
pubmed-meshheading:10049066-Humans,
pubmed-meshheading:10049066-Mice,
pubmed-meshheading:10049066-Stromal Cells,
pubmed-meshheading:10049066-Time Factors
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pubmed:year |
1999
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pubmed:articleTitle |
The type of stromal feeder used in limiting dilution assays influences frequency and maintenance assessment of human long-term culture initiating cells.
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pubmed:affiliation |
Department of Medicine, and Stem Cell Biology Program, University of Minnesota, Minneapolis 55455, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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