In Vitro proliferation of primitive hemopoietic stem cells supported by stromal cells: Evidence for the presence of a mechanism(s) other than that involving c-kit receptor and its ligand

Hiroaki Kodama, Makoto Nose, Yuji Yamaguchi, Jun Ichi Tsunoda, Toshio Suda, Satomi Nishikawa, Shin Ichi Nishikawa

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Abstract

The preadipose cell line, PA6, can support long-term hemopoiesis. Frequency of the hemopoietic stem cells capable of sustaining hemopoiesis in cocultures of bone marrow cells and PA6 cells for 6 wk was 1/5.3 × 104 bone marrow cells. In the group of dishes into which bone marrow cells had been inoculated at 2.5 × 104 cells/dish, 3 of 19 dishes (16%) contained stem cells capable of reconstituting erythropoiesis of WBB6F1-W/Wv mice, indicating that PA6 cells can support the proliferation of primitive hemopoietic stem cells. When the cocultures were treated with an antagonistic anti-c-kit monoclonal antibody, ACK2, only a small number of day 12 spleen colony-forming units survived; and hemopoiesis was severely reduced. However, when the cocultures were continued with antibody-free medium, hemopoiesis dramatically recovered. To examine the proliferative properties of the ACK2-resistant stem cells, we developed a colony assay system by modifying our coculture system. Sequential observations of the development of individual colonies and their disappearance demonstrated that the stem cells having higher proliferative capacity preferentially survive the ACK2 treatment. Furthermore, cells of subclones of the PA6 clone that were incapable of supporting long-term hemopoiesis expressed mRNA for the c-kit ligand. These results suggest that a mechanism(s) other than that involving c-kit receptor and its ligand plays an important role in the survival and proliferation of primitive hemopoietic stem cells.

Original languageEnglish
Pages (from-to)351-361
Number of pages11
JournalJournal of Experimental Medicine
Volume176
Issue number2
Publication statusPublished - 1992 Aug 1

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Proto-Oncogene Proteins c-kit
Stromal Cells
Stem Cells
Ligands
Coculture Techniques
Bone Marrow Cells
Stem Cell Factor
Erythropoiesis
In Vitro Techniques
Spleen
Clone Cells
Monoclonal Antibodies
Cell Line
Messenger RNA
Antibodies

ASJC Scopus subject areas

  • Immunology

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In Vitro proliferation of primitive hemopoietic stem cells supported by stromal cells : Evidence for the presence of a mechanism(s) other than that involving c-kit receptor and its ligand. / Kodama, Hiroaki; Nose, Makoto; Yamaguchi, Yuji; Tsunoda, Jun Ichi; Suda, Toshio; Nishikawa, Satomi; Nishikawa, Shin Ichi.

In: Journal of Experimental Medicine, Vol. 176, No. 2, 01.08.1992, p. 351-361.

Research output: Contribution to journalArticle

Kodama, Hiroaki ; Nose, Makoto ; Yamaguchi, Yuji ; Tsunoda, Jun Ichi ; Suda, Toshio ; Nishikawa, Satomi ; Nishikawa, Shin Ichi. / In Vitro proliferation of primitive hemopoietic stem cells supported by stromal cells : Evidence for the presence of a mechanism(s) other than that involving c-kit receptor and its ligand. In: Journal of Experimental Medicine. 1992 ; Vol. 176, No. 2. pp. 351-361.
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