Cell-stoma interactions in haemopoiesis studied by immunocytochemistry and in situ hybridisation in long-term cultures and trephine biopsies in human bone marrow
Cell-stoma interactions in haemopoiesis studied by immunocytochemistry and in situ hybridisation in long-term cultures and trephine biopsies in human bone marrow
This study was undertaken to characterise the stroma which supports haemopoietic cells in bone marrow. Human long-term bone marrow cultures (hLTBMC) were grown, in which stromal elements predominate and localised haemopoietic foci form. Trephine biopsies (BMT) were also studied, representing samples of intact bone marrow. Biopsies and samples for culture were obtained from patients undergoing haematological investigations for a variety of disorders; the bone marrow studied therefore included normal and pathological material.
Cellular and matrix components of hLTBMC and BMT were investigated using immunocytochemistry and messenger RNA in situ hybridisation (ISH). Immunostaining of hLTBMC demonstrated the presence of a variety of fibroblastic cells, including ASMA+ve polygonal cells and NGFR+ve dendritic cells, with adherent populations of round and flattened macrophages. Organised capillary-like structures also formed within hLTBMC stroma. A variety of adhesion molecules was expressed by stromal cells matrix and haemopoietic cells but evidence of pairing with known ligands was absent in each case, suggesting that alternative ligands may exist for the molecules demonstrated.
In situ hybridisation was undertaken to demonstrate mRNA species encoding the haemopoietic growth factors IL-3, GM-CSF, G-CSF and M-CSF. Weak expression of GM-CSF and M-CSF by stromal fibroblasts was demonstrated, but the other mRNAs were undetectable by this technique. Stronger expression of M-CSF by monocytes and macrophages was found, suggesting that an autocrine mechanism may contribute to monocyte/macrophage differentiation.
Immunostaining and ISH methods developed for this study were also used to investigate the hyperplastic bone marrow response seen in patients with Hodgkin's disease. Production of GM-CSF, G-CSF and M-CSF by Reed-Sternberg cells and accompanying reactive cells was shown but there was marked variation between individuals and no correlation was found with the extent of bone marrow reaction present.
University of Southampton
Wilkins, Bridget Sally
1192664a-9636-4d48-80e8-6eda9ce8f423
1996
Wilkins, Bridget Sally
1192664a-9636-4d48-80e8-6eda9ce8f423
Wilkins, Bridget Sally
(1996)
Cell-stoma interactions in haemopoiesis studied by immunocytochemistry and in situ hybridisation in long-term cultures and trephine biopsies in human bone marrow.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
This study was undertaken to characterise the stroma which supports haemopoietic cells in bone marrow. Human long-term bone marrow cultures (hLTBMC) were grown, in which stromal elements predominate and localised haemopoietic foci form. Trephine biopsies (BMT) were also studied, representing samples of intact bone marrow. Biopsies and samples for culture were obtained from patients undergoing haematological investigations for a variety of disorders; the bone marrow studied therefore included normal and pathological material.
Cellular and matrix components of hLTBMC and BMT were investigated using immunocytochemistry and messenger RNA in situ hybridisation (ISH). Immunostaining of hLTBMC demonstrated the presence of a variety of fibroblastic cells, including ASMA+ve polygonal cells and NGFR+ve dendritic cells, with adherent populations of round and flattened macrophages. Organised capillary-like structures also formed within hLTBMC stroma. A variety of adhesion molecules was expressed by stromal cells matrix and haemopoietic cells but evidence of pairing with known ligands was absent in each case, suggesting that alternative ligands may exist for the molecules demonstrated.
In situ hybridisation was undertaken to demonstrate mRNA species encoding the haemopoietic growth factors IL-3, GM-CSF, G-CSF and M-CSF. Weak expression of GM-CSF and M-CSF by stromal fibroblasts was demonstrated, but the other mRNAs were undetectable by this technique. Stronger expression of M-CSF by monocytes and macrophages was found, suggesting that an autocrine mechanism may contribute to monocyte/macrophage differentiation.
Immunostaining and ISH methods developed for this study were also used to investigate the hyperplastic bone marrow response seen in patients with Hodgkin's disease. Production of GM-CSF, G-CSF and M-CSF by Reed-Sternberg cells and accompanying reactive cells was shown but there was marked variation between individuals and no correlation was found with the extent of bone marrow reaction present.
This record has no associated files available for download.
More information
Published date: 1996
Identifiers
Local EPrints ID: 459500
URI: http://eprints.soton.ac.uk/id/eprint/459500
PURE UUID: f50f2e91-b893-42f7-956c-57515795112f
Catalogue record
Date deposited: 04 Jul 2022 17:13
Last modified: 23 Jul 2022 00:30
Export record
Contributors
Author:
Bridget Sally Wilkins
Download statistics
Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.
View more statistics