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Diffusion Chamber Culture pp 124–135Cite as

Stimulating Factor(s) of Hematopoiesis in the Bone Marrow of Mice Administered Hydroxyurea. Effects of Diffusion Chamber Progenitor Cells (DCPC) and Observations on Pluripotent Stem Cells (CFU-S)

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Abstract

A study of hematopoiesis in Sabra mice was carried out soon after hydroxyurea (HU) administration. Three hours after administration of a single intra-peritoneal injection of HU (1000 mg/kg), 50% of the nucleated bone marrow cells per femur were killed. HU-treated bone marrow was inoculated into peritoneal DC in non-irradiated mice. In additional experiments in which the double diffusion chamber (DDC) technique was used, HU-treated bone marrow was inoculated into the regulator compartment (RC) and normal bone marrow into the test compartment (TC). The DDC were implanted in irradiated hosts and cultured for 7 days. In control experiments, normal bone marrow was inoculated into both RC and TC. The results obtained with DC and DDC demonstrate the presence of stimulating factors of diffusion chamber progenitor cells (DCPC) in the bone marrow of mice 3 h following HU administration. The occurrence of a stimulating factor(s) of granulopoiesis was manifested by (1) a higher granulocyte/macrophage (G/M) ratio in single DC containing HU-treated bone marrow than in control cultures and (2) a higher G/M ratio in both compartments of the DDC in experimental groups than in controls. The stimulating factor of granulopoiesis present in HU-treated bone marrow crossed the Millipore filter (0.2 μm pore size) separating the compartments of the DDC as manifested by a high G/M ratio in both RC and TC.

The cell yield in the RC of the DDC was significantly higher than that of the TC in 7 day cultures. This finding is indicative of a process of self-replication of DCPC and/or of mobilization of more primitive DCPC (“pre-DCPC”) following HU administration.

The spleen colony assay was carried out in groups of lethally irradiated mice which received intravenously: (a) normal murine bone marrow cells, (b) bone marrow of donors which were administered a single dose of HU 3 h prior to marrow collection, and (c) bone marrow of mice administered HU 9 h prior to death. The results with spleen colonies indicate that 3 h following HU administration to mice, the pluripotent stem cells are not damaged, apparently because the majority of CFU-S are noncycling cells in Sabra mice. However, 9 h following HU administration, the resulting hypocellular bone marrow appears to be 2 times more concentrated in CFU-S than the normal marrow.

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Authors
  1. Z. Ben-Ishay
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  2. S. Sharon
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  3. A. Shorr
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Editor information

Editors and Affiliations

  1. Medical Department, Brookhaven National Laboratory, Upton, N.Y., USA

    Eugene P. Cronkite MD  & Arland L. Carsten PhD  & 

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© 1980 Springer-Verlag Berlin Heidelberg

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Ben-Ishay, Z., Sharon, S., Shorr, A. (1980). Stimulating Factor(s) of Hematopoiesis in the Bone Marrow of Mice Administered Hydroxyurea. Effects of Diffusion Chamber Progenitor Cells (DCPC) and Observations on Pluripotent Stem Cells (CFU-S). In: Cronkite, E.P., Carsten, A.L. (eds) Diffusion Chamber Culture. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-67644-4_12

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  • DOI: https://doi.org/10.1007/978-3-642-67644-4_12

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-540-10064-5

  • Online ISBN: 978-3-642-67644-4

  • eBook Packages: Springer Book Archive

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