A positive effect of p21(cip1/waf1) in the colony formation from murine myeloid progenitor cells as assessed by retroviral-mediated gene transfer

Stephen E. Braun, Charlie Mantel, Marc Rosenthal, Scott Cooper, Lisa Liu, Kent Robertson, Robert Hromas, Hal Broxmeyer

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

p21(cip1/waf1) is a cyclin dependent kinase inhibitor. We have previously reported stimulation of p21(cip1/waf1) by steel factor and GM-CSF in a factor dependent cell line and of p21(cip1/waf1) involvement in hematopoiesis in vivo in p21(cip1/waf1) gene knockout (-/-) mice. To further assess a role for increased p21(cip1/waf1) in hematopoietic progenitor cells, we developed the retroviral vector L(p21(cip1))SN to transcriptionally regulate p21(cip1/waf1) from the Mo-MLV LTR. L(p21(cip1))SN and the control vector LXSN were used to transduce murine bone marrow progenitor cells from p21(cip1/waf1) (-/-) and littermate control (+/+) mice, as well as from other mouse strains. Hematopoietic colony formation by transduced cells was assessed in semi-solid culture medium with multiple growth factors. Myeloid colony formation by bone marrow cells from p21(cip1/waf1) (-/-) mice was significantly lower than that by (+/+) mouse cells. Transduction of cells with LXSN had no effect on colony formation; however, (-/-) cells transduced with L(p21(cip1))SN formed significantly greater numbers of colonies than either LXSN-transduced (-/-) or (+/+) cells. Moreover, L(p21(cip1))SN- transduced (+/+) cells formed significantly more colonies than LXSN- transduced (+/+) cells. Increased cloning efficiency of progenitors from normal strains of mice induced by L(p21(cip1))SN compared to LXSN-transduced cells was seen whether unseparated or highly purified populations of Scal+ Lin- marrow cells were used. Gene transfer of L(p21(cip1))SN increased the size and number of cells per colony, as well as the number of colonies compared to LXSN gene transfer. No colonies grew from non-transduced, LXSN- , or L(p21(cip1))SN-transduced cells when no growth factors were added to the cultures. These results document the positive effect of p21(cip1/waf1) in the proliferation and/or differentiation of the murine myeloid progenitor cells that lead to colony formation.

Original languageEnglish
Pages (from-to)138-148
Number of pages11
JournalBlood Cells, Molecules and Diseases
Volume24
Issue number2
DOIs
StatePublished - Jun 1998

Fingerprint

Myeloid Progenitor Cells
Genes
Bone Marrow Cells
Intercellular Signaling Peptides and Proteins
Gene Knockout Techniques
Stem Cell Factor
Cyclin-Dependent Kinases
Hematopoiesis
Granulocyte-Macrophage Colony-Stimulating Factor
Hematopoietic Stem Cells
Cell Size
Knockout Mice
Culture Media
Organism Cloning
Stem Cells
Cell Count
Bone Marrow

Keywords

  • Gene transfer
  • Hematopoietic progenitor cells
  • p21(cip1/waf1)
  • p21(cip1/waf1) knockout mice

ASJC Scopus subject areas

  • Molecular Biology
  • Molecular Medicine
  • Hematology

Cite this

A positive effect of p21(cip1/waf1) in the colony formation from murine myeloid progenitor cells as assessed by retroviral-mediated gene transfer. / Braun, Stephen E.; Mantel, Charlie; Rosenthal, Marc; Cooper, Scott; Liu, Lisa; Robertson, Kent; Hromas, Robert; Broxmeyer, Hal.

In: Blood Cells, Molecules and Diseases, Vol. 24, No. 2, 06.1998, p. 138-148.

Research output: Contribution to journalArticle

Braun, Stephen E. ; Mantel, Charlie ; Rosenthal, Marc ; Cooper, Scott ; Liu, Lisa ; Robertson, Kent ; Hromas, Robert ; Broxmeyer, Hal. / A positive effect of p21(cip1/waf1) in the colony formation from murine myeloid progenitor cells as assessed by retroviral-mediated gene transfer. In: Blood Cells, Molecules and Diseases. 1998 ; Vol. 24, No. 2. pp. 138-148.
@article{3fd2f9e3a8d046c0bfba859a940f7583,
title = "A positive effect of p21(cip1/waf1) in the colony formation from murine myeloid progenitor cells as assessed by retroviral-mediated gene transfer",
abstract = "p21(cip1/waf1) is a cyclin dependent kinase inhibitor. We have previously reported stimulation of p21(cip1/waf1) by steel factor and GM-CSF in a factor dependent cell line and of p21(cip1/waf1) involvement in hematopoiesis in vivo in p21(cip1/waf1) gene knockout (-/-) mice. To further assess a role for increased p21(cip1/waf1) in hematopoietic progenitor cells, we developed the retroviral vector L(p21(cip1))SN to transcriptionally regulate p21(cip1/waf1) from the Mo-MLV LTR. L(p21(cip1))SN and the control vector LXSN were used to transduce murine bone marrow progenitor cells from p21(cip1/waf1) (-/-) and littermate control (+/+) mice, as well as from other mouse strains. Hematopoietic colony formation by transduced cells was assessed in semi-solid culture medium with multiple growth factors. Myeloid colony formation by bone marrow cells from p21(cip1/waf1) (-/-) mice was significantly lower than that by (+/+) mouse cells. Transduction of cells with LXSN had no effect on colony formation; however, (-/-) cells transduced with L(p21(cip1))SN formed significantly greater numbers of colonies than either LXSN-transduced (-/-) or (+/+) cells. Moreover, L(p21(cip1))SN- transduced (+/+) cells formed significantly more colonies than LXSN- transduced (+/+) cells. Increased cloning efficiency of progenitors from normal strains of mice induced by L(p21(cip1))SN compared to LXSN-transduced cells was seen whether unseparated or highly purified populations of Scal+ Lin- marrow cells were used. Gene transfer of L(p21(cip1))SN increased the size and number of cells per colony, as well as the number of colonies compared to LXSN gene transfer. No colonies grew from non-transduced, LXSN- , or L(p21(cip1))SN-transduced cells when no growth factors were added to the cultures. These results document the positive effect of p21(cip1/waf1) in the proliferation and/or differentiation of the murine myeloid progenitor cells that lead to colony formation.",
keywords = "Gene transfer, Hematopoietic progenitor cells, p21(cip1/waf1), p21(cip1/waf1) knockout mice",
author = "Braun, {Stephen E.} and Charlie Mantel and Marc Rosenthal and Scott Cooper and Lisa Liu and Kent Robertson and Robert Hromas and Hal Broxmeyer",
year = "1998",
month = "6",
doi = "10.1006/bcmd.1998.0181",
language = "English",
volume = "24",
pages = "138--148",
journal = "Blood Cells, Molecules, and Diseases",
issn = "1079-9796",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - A positive effect of p21(cip1/waf1) in the colony formation from murine myeloid progenitor cells as assessed by retroviral-mediated gene transfer

AU - Braun, Stephen E.

AU - Mantel, Charlie

AU - Rosenthal, Marc

AU - Cooper, Scott

AU - Liu, Lisa

AU - Robertson, Kent

AU - Hromas, Robert

AU - Broxmeyer, Hal

PY - 1998/6

Y1 - 1998/6

N2 - p21(cip1/waf1) is a cyclin dependent kinase inhibitor. We have previously reported stimulation of p21(cip1/waf1) by steel factor and GM-CSF in a factor dependent cell line and of p21(cip1/waf1) involvement in hematopoiesis in vivo in p21(cip1/waf1) gene knockout (-/-) mice. To further assess a role for increased p21(cip1/waf1) in hematopoietic progenitor cells, we developed the retroviral vector L(p21(cip1))SN to transcriptionally regulate p21(cip1/waf1) from the Mo-MLV LTR. L(p21(cip1))SN and the control vector LXSN were used to transduce murine bone marrow progenitor cells from p21(cip1/waf1) (-/-) and littermate control (+/+) mice, as well as from other mouse strains. Hematopoietic colony formation by transduced cells was assessed in semi-solid culture medium with multiple growth factors. Myeloid colony formation by bone marrow cells from p21(cip1/waf1) (-/-) mice was significantly lower than that by (+/+) mouse cells. Transduction of cells with LXSN had no effect on colony formation; however, (-/-) cells transduced with L(p21(cip1))SN formed significantly greater numbers of colonies than either LXSN-transduced (-/-) or (+/+) cells. Moreover, L(p21(cip1))SN- transduced (+/+) cells formed significantly more colonies than LXSN- transduced (+/+) cells. Increased cloning efficiency of progenitors from normal strains of mice induced by L(p21(cip1))SN compared to LXSN-transduced cells was seen whether unseparated or highly purified populations of Scal+ Lin- marrow cells were used. Gene transfer of L(p21(cip1))SN increased the size and number of cells per colony, as well as the number of colonies compared to LXSN gene transfer. No colonies grew from non-transduced, LXSN- , or L(p21(cip1))SN-transduced cells when no growth factors were added to the cultures. These results document the positive effect of p21(cip1/waf1) in the proliferation and/or differentiation of the murine myeloid progenitor cells that lead to colony formation.

AB - p21(cip1/waf1) is a cyclin dependent kinase inhibitor. We have previously reported stimulation of p21(cip1/waf1) by steel factor and GM-CSF in a factor dependent cell line and of p21(cip1/waf1) involvement in hematopoiesis in vivo in p21(cip1/waf1) gene knockout (-/-) mice. To further assess a role for increased p21(cip1/waf1) in hematopoietic progenitor cells, we developed the retroviral vector L(p21(cip1))SN to transcriptionally regulate p21(cip1/waf1) from the Mo-MLV LTR. L(p21(cip1))SN and the control vector LXSN were used to transduce murine bone marrow progenitor cells from p21(cip1/waf1) (-/-) and littermate control (+/+) mice, as well as from other mouse strains. Hematopoietic colony formation by transduced cells was assessed in semi-solid culture medium with multiple growth factors. Myeloid colony formation by bone marrow cells from p21(cip1/waf1) (-/-) mice was significantly lower than that by (+/+) mouse cells. Transduction of cells with LXSN had no effect on colony formation; however, (-/-) cells transduced with L(p21(cip1))SN formed significantly greater numbers of colonies than either LXSN-transduced (-/-) or (+/+) cells. Moreover, L(p21(cip1))SN- transduced (+/+) cells formed significantly more colonies than LXSN- transduced (+/+) cells. Increased cloning efficiency of progenitors from normal strains of mice induced by L(p21(cip1))SN compared to LXSN-transduced cells was seen whether unseparated or highly purified populations of Scal+ Lin- marrow cells were used. Gene transfer of L(p21(cip1))SN increased the size and number of cells per colony, as well as the number of colonies compared to LXSN gene transfer. No colonies grew from non-transduced, LXSN- , or L(p21(cip1))SN-transduced cells when no growth factors were added to the cultures. These results document the positive effect of p21(cip1/waf1) in the proliferation and/or differentiation of the murine myeloid progenitor cells that lead to colony formation.

KW - Gene transfer

KW - Hematopoietic progenitor cells

KW - p21(cip1/waf1)

KW - p21(cip1/waf1) knockout mice

UR - http://www.scopus.com/inward/record.url?scp=0032081244&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032081244&partnerID=8YFLogxK

U2 - 10.1006/bcmd.1998.0181

DO - 10.1006/bcmd.1998.0181

M3 - Article

C2 - 9628851

AN - SCOPUS:0032081244

VL - 24

SP - 138

EP - 148

JO - Blood Cells, Molecules, and Diseases

JF - Blood Cells, Molecules, and Diseases

SN - 1079-9796

IS - 2

ER -