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Terminal differentiation of murine erythroleukemia cells: physical stabilization of end- stage cells

An important limitation in the use of the murine erythroleukenia (MEL) cell system as an in vitro system for the study of terminal erythroid differentiation has been the inability to produce significant numbers of cells which represent the end-point of the pathway in vitro. We show here that a major...

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Detalles Bibliográficos
Autores principales: Volloch, V, Housman, D
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 1982
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2112864/
https://www.ncbi.nlm.nih.gov/pubmed/6954153
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author Volloch, V
Housman, D
author_facet Volloch, V
Housman, D
author_sort Volloch, V
collection PubMed
description An important limitation in the use of the murine erythroleukenia (MEL) cell system as an in vitro system for the study of terminal erythroid differentiation has been the inability to produce significant numbers of cells which represent the end-point of the pathway in vitro. We show here that a major reason for the failure to observe end-stage cells in vitro is that such cells are physically unstable under the standard culture conditions used for MEL cell differentiation. Modification of these culture conditions by the addition of either bovine serum albumin or Ficoll leads to physical stabilization of end-stage cells. Under such culture conditions, uniform cultures of terminally differentiated MEL cells with morphological characteristics similar to those of normal mouse orthochromatophilic erythroblasts and reticulocytes are observed. Examination of physical and biochemical parameters of these cell populations give values which are similar to values characteristic of mouse reticulocytes. A physically stabilized MEL cell shows a narrow cell volume distribution with an average value of approximately 100 mum(3), similar to the cell volume distribution observed for mouse reticulocytes, while a typical MEL cell culture treated with DMSO but without a stabilizing agent exhibits a broader, more heterogeneous cell volume distribution with an average value of approximately 500 mum(3). Globin mRNA levels and levels of globin synthesis reach values almost equal to those in mouse reticulocytes in cultures of physically stabilized MEL cells while differentiating cultures not treated with a stabilizing agent reach substantially lower values for these parameters. We suggest that the ability to produce populations of MEL cells which undergo complete terminal erythroid differentiation in vitro will allow the analysis of the molecular mechanisms which control the terminal stages of the erythroid differentiation process.
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spelling pubmed-21128642008-05-01 Terminal differentiation of murine erythroleukemia cells: physical stabilization of end- stage cells Volloch, V Housman, D J Cell Biol Articles An important limitation in the use of the murine erythroleukenia (MEL) cell system as an in vitro system for the study of terminal erythroid differentiation has been the inability to produce significant numbers of cells which represent the end-point of the pathway in vitro. We show here that a major reason for the failure to observe end-stage cells in vitro is that such cells are physically unstable under the standard culture conditions used for MEL cell differentiation. Modification of these culture conditions by the addition of either bovine serum albumin or Ficoll leads to physical stabilization of end-stage cells. Under such culture conditions, uniform cultures of terminally differentiated MEL cells with morphological characteristics similar to those of normal mouse orthochromatophilic erythroblasts and reticulocytes are observed. Examination of physical and biochemical parameters of these cell populations give values which are similar to values characteristic of mouse reticulocytes. A physically stabilized MEL cell shows a narrow cell volume distribution with an average value of approximately 100 mum(3), similar to the cell volume distribution observed for mouse reticulocytes, while a typical MEL cell culture treated with DMSO but without a stabilizing agent exhibits a broader, more heterogeneous cell volume distribution with an average value of approximately 500 mum(3). Globin mRNA levels and levels of globin synthesis reach values almost equal to those in mouse reticulocytes in cultures of physically stabilized MEL cells while differentiating cultures not treated with a stabilizing agent reach substantially lower values for these parameters. We suggest that the ability to produce populations of MEL cells which undergo complete terminal erythroid differentiation in vitro will allow the analysis of the molecular mechanisms which control the terminal stages of the erythroid differentiation process. The Rockefeller University Press 1982-05-01 /pmc/articles/PMC2112864/ /pubmed/6954153 Text en This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Articles
Volloch, V
Housman, D
Terminal differentiation of murine erythroleukemia cells: physical stabilization of end- stage cells
title Terminal differentiation of murine erythroleukemia cells: physical stabilization of end- stage cells
title_full Terminal differentiation of murine erythroleukemia cells: physical stabilization of end- stage cells
title_fullStr Terminal differentiation of murine erythroleukemia cells: physical stabilization of end- stage cells
title_full_unstemmed Terminal differentiation of murine erythroleukemia cells: physical stabilization of end- stage cells
title_short Terminal differentiation of murine erythroleukemia cells: physical stabilization of end- stage cells
title_sort terminal differentiation of murine erythroleukemia cells: physical stabilization of end- stage cells
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2112864/
https://www.ncbi.nlm.nih.gov/pubmed/6954153
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AT housmand terminaldifferentiationofmurineerythroleukemiacellsphysicalstabilizationofendstagecells