Hemoglobin synthesis in somatic cell hybrids

globin gene expression in hybrids between mouse erythroleukemia and human marrow cells or fibroblasts

A. Deisseroth, Robert D. Burk, D. Picciano, W. F. Anderson, A. Nienhuis, J. Minna

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Somatic cell hybrids were generated by fusion of mouse erythroleukemia cells either to mouse L cells (B82), human fibroblasts (W1-18 VA2), or human marrow fractions enriched in erythroblasts. The hybrid cells were examined for globin gene expression by benzidine staining to detect cytoplasmic hemoglobin, and by molecular hybridization of cellular RNA to globin complementary DNA (cDNA) to detect globin messenger RNA (mRNA). The fibroblast (human or mouse) x erythroleukemia cell hybrids grown in monolayer retained most of the chromosomes of each parent. Neither cytoplasmic hemoglobin nor globin mRNA was detected in dimethylsulfoxide treated fibroblast x erythroleukemia hybrid cells, indicating extinction of hemoglobin synthesis prior to the formation of cytoplasmic mRNA. The human marrow x mouse erythroleukemia hybrid cells grown in suspension culture contained only a few human chromosomes and exhibited low levels of hemoglobin synthesis which were amplified by 2% dimethylsulfoxide. Mouse (but not human) globin mRNA was demonstrated in these hybrid cells. The results suggest that somatic cell hybrids may be useful in searching for genetic factors which regulate activity of the globulin genes.

Original languageEnglish (US)
Pages (from-to)1102-1106
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume72
Issue number3
DOIs
StatePublished - 1975
Externally publishedYes

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Leukemia, Erythroblastic, Acute
Globins
Hybrid Cells
Hemoglobins
Fibroblasts
Bone Marrow
Gene Expression
Messenger RNA
Dimethyl Sulfoxide
Erythroblasts
Globulins
Human Chromosomes
Suspensions
Complementary DNA
Chromosomes
RNA
Staining and Labeling
Genes

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Hemoglobin synthesis in somatic cell hybrids : globin gene expression in hybrids between mouse erythroleukemia and human marrow cells or fibroblasts. / Deisseroth, A.; Burk, Robert D.; Picciano, D.; Anderson, W. F.; Nienhuis, A.; Minna, J.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 72, No. 3, 1975, p. 1102-1106.

Research output: Contribution to journalArticle

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AU - Anderson, W. F.

AU - Nienhuis, A.

AU - Minna, J.

PY - 1975

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N2 - Somatic cell hybrids were generated by fusion of mouse erythroleukemia cells either to mouse L cells (B82), human fibroblasts (W1-18 VA2), or human marrow fractions enriched in erythroblasts. The hybrid cells were examined for globin gene expression by benzidine staining to detect cytoplasmic hemoglobin, and by molecular hybridization of cellular RNA to globin complementary DNA (cDNA) to detect globin messenger RNA (mRNA). The fibroblast (human or mouse) x erythroleukemia cell hybrids grown in monolayer retained most of the chromosomes of each parent. Neither cytoplasmic hemoglobin nor globin mRNA was detected in dimethylsulfoxide treated fibroblast x erythroleukemia hybrid cells, indicating extinction of hemoglobin synthesis prior to the formation of cytoplasmic mRNA. The human marrow x mouse erythroleukemia hybrid cells grown in suspension culture contained only a few human chromosomes and exhibited low levels of hemoglobin synthesis which were amplified by 2% dimethylsulfoxide. Mouse (but not human) globin mRNA was demonstrated in these hybrid cells. The results suggest that somatic cell hybrids may be useful in searching for genetic factors which regulate activity of the globulin genes.

AB - Somatic cell hybrids were generated by fusion of mouse erythroleukemia cells either to mouse L cells (B82), human fibroblasts (W1-18 VA2), or human marrow fractions enriched in erythroblasts. The hybrid cells were examined for globin gene expression by benzidine staining to detect cytoplasmic hemoglobin, and by molecular hybridization of cellular RNA to globin complementary DNA (cDNA) to detect globin messenger RNA (mRNA). The fibroblast (human or mouse) x erythroleukemia cell hybrids grown in monolayer retained most of the chromosomes of each parent. Neither cytoplasmic hemoglobin nor globin mRNA was detected in dimethylsulfoxide treated fibroblast x erythroleukemia hybrid cells, indicating extinction of hemoglobin synthesis prior to the formation of cytoplasmic mRNA. The human marrow x mouse erythroleukemia hybrid cells grown in suspension culture contained only a few human chromosomes and exhibited low levels of hemoglobin synthesis which were amplified by 2% dimethylsulfoxide. Mouse (but not human) globin mRNA was demonstrated in these hybrid cells. The results suggest that somatic cell hybrids may be useful in searching for genetic factors which regulate activity of the globulin genes.

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