Measurement of long-term culture initiating cells (LTC-ICs) using limiting dilution: comparison of endpoints and stromal support.

2.50
Hdl Handle:
http://hdl.handle.net/10541/94884
Title:
Measurement of long-term culture initiating cells (LTC-ICs) using limiting dilution: comparison of endpoints and stromal support.
Authors:
Weaver, A; Ryder, W David J; Testa, Nydia G
Abstract:
Although much progress has been made in defining primitive cell phenotypes using flow cytometry and clonogenic assays, the direct study of marrow repopulating cells remains elusive. Long-term culture initiating cells (LTC-ICs) are arguably the most primitive human hematopoietic cells detectable by in vitro functional assays. Two endpoints have been reported for scoring LTC-ICs in limiting dilution assays. The first endpoint described was the generation of colony forming cells (CFCs) after 5 to 8 weeks of culture. An alternative method for scoring the LTC-IC assay is to identify cobblestone area forming cells. In the present study, estimations of LTC-IC frequency were made by measuring both endpoints and by comparing LTC-IC frequencies measured using limiting dilution assays of normal human bone marrow stroma with the measurements for murine bone marrow stromal cell line M2-10B4. For assays established on normal human bone marrow stroma, there was an equivalence between the two endpoint measures. Likewise, there was an equivalence between the two types of stroma when scoring CFC generation after 5 weeks. However, a consistently higher frequency of LTC-ICs was estimated when scoring cobblestone areas compared with that found when scoring CFC generation on the M2-10B4 stroma (p < 0.0001). Although the murine bone marrow stromal cell line M2-10B4 remains a very useful and consistently reliable alternative to normal human bone marrow stroma, these data indicate that the LTC-IC populations defined by scoring cobblestone areas or by measuring the generation of CFCs on this cell line are, in contrast to those measured using bone marrow stroma, not identical.
Affiliation:
Department of Experimental Haematology, Paterson Institute for Cancer Research, United Kingdom.
Citation:
Measurement of long-term culture initiating cells (LTC-ICs) using limiting dilution: comparison of endpoints and stromal support. 1997, 25 (13):1333-8 Exp. Hematol.
Journal:
Experimental Hematology
Issue Date:
Dec-1997
URI:
http://hdl.handle.net/10541/94884
PubMed ID:
9406992
Type:
Article
Language:
en
ISSN:
0301-472X
Appears in Collections:
All Christie Publications ; All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorWeaver, Aen
dc.contributor.authorRyder, W David Jen
dc.contributor.authorTesta, Nydia Gen
dc.date.accessioned2010-03-24T14:30:52Z-
dc.date.available2010-03-24T14:30:52Z-
dc.date.issued1997-12-
dc.identifier.citationMeasurement of long-term culture initiating cells (LTC-ICs) using limiting dilution: comparison of endpoints and stromal support. 1997, 25 (13):1333-8 Exp. Hematol.en
dc.identifier.issn0301-472X-
dc.identifier.pmid9406992-
dc.identifier.urihttp://hdl.handle.net/10541/94884-
dc.description.abstractAlthough much progress has been made in defining primitive cell phenotypes using flow cytometry and clonogenic assays, the direct study of marrow repopulating cells remains elusive. Long-term culture initiating cells (LTC-ICs) are arguably the most primitive human hematopoietic cells detectable by in vitro functional assays. Two endpoints have been reported for scoring LTC-ICs in limiting dilution assays. The first endpoint described was the generation of colony forming cells (CFCs) after 5 to 8 weeks of culture. An alternative method for scoring the LTC-IC assay is to identify cobblestone area forming cells. In the present study, estimations of LTC-IC frequency were made by measuring both endpoints and by comparing LTC-IC frequencies measured using limiting dilution assays of normal human bone marrow stroma with the measurements for murine bone marrow stromal cell line M2-10B4. For assays established on normal human bone marrow stroma, there was an equivalence between the two endpoint measures. Likewise, there was an equivalence between the two types of stroma when scoring CFC generation after 5 weeks. However, a consistently higher frequency of LTC-ICs was estimated when scoring cobblestone areas compared with that found when scoring CFC generation on the M2-10B4 stroma (p < 0.0001). Although the murine bone marrow stromal cell line M2-10B4 remains a very useful and consistently reliable alternative to normal human bone marrow stroma, these data indicate that the LTC-IC populations defined by scoring cobblestone areas or by measuring the generation of CFCs on this cell line are, in contrast to those measured using bone marrow stroma, not identical.en
dc.language.isoenen
dc.subjectHaematopoietic Stem Cell Mobilisationen
dc.subjectHaematopoietic Stem Cellsen
dc.subjectOvarian Canceren
dc.subject.meshAnimals-
dc.subject.meshBone Marrow-
dc.subject.meshCell Culture Techniques-
dc.subject.meshColony-Forming Units Assay-
dc.subject.meshFemale-
dc.subject.meshFlow Cytometry-
dc.subject.meshGranulocyte-Macrophage Colony-Stimulating Factor-
dc.subject.meshHematopoietic Stem Cell Mobilization-
dc.subject.meshHematopoietic Stem Cells-
dc.subject.meshHumans-
dc.subject.meshIndicator Dilution Techniques-
dc.subject.meshOvarian Neoplasms-
dc.subject.meshStromal Cells-
dc.titleMeasurement of long-term culture initiating cells (LTC-ICs) using limiting dilution: comparison of endpoints and stromal support.en
dc.typeArticleen
dc.contributor.departmentDepartment of Experimental Haematology, Paterson Institute for Cancer Research, United Kingdom.en
dc.identifier.journalExperimental Hematologyen
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