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Agar Technique for the Cultivation In Vitro of Bone-Marrow Colonies

Abstract

In solid-state agar cultures certain haemopoietic cells proliferate and form discrete colonies of 200 - 4000 cells. Colony formation is dependent on stimulation by the colony-stimulating factor, and this is achieved by (1) the use of a cell feeder layer, (2) the addition of conditioned medium, or (3) the addition of human or mouse serum or urine containing the factor. All colonies initially contain granulocytic cells which differentiate from myeloblasts to polymorphs as colony growth proceeds. Later colonies develop a second population of phagocytic mononuclear cells (macrophages). The colony-forming-system is simple, readily quantitated and highly reproducible. Linear dose responses occur between the dose of colony-stimulating factor and the number and size of colonies developing from a standard number of bone-marrow cells. In-vitro colony formation has been achieved with haemopoietic cells of the following species: mouse, rat, hamster, guinea pig, rabbit and human. In the adult mouse, colony-forming cells are located in the bone marrow, spleen and blood and in the embryo, in the yolk sac, liver and spleen. The colony-forming cell appears to be an early member of the granulocytic series. The colony-forming system has been used as a quantitative assay system: (1) to assay levels of colony-stimulating factor in  More>>
Authors:
Metcalf, D. [1] 
  1. Walter and Eliza Hall Institute, Royal Melbourne Hospital, Melbourne, VIC (Australia)
Publication Date:
Jul 15, 1969
Product Type:
Conference
Resource Relation:
Conference: Panel on Current Problems of Bone-Marrow Cell Transplantation with Special Emphasis on Conservation and Culture, Moscow (Russian Federation), 22-26 Jul 1968; Other Information: 22 refs.; Related Information: In: Bone-Marrow Conservation, Culture and Transplantation. Proceedings of a Panel on Current Problems of Bone-Marrow Cell Transplantation with Special Emphasis on Conservation and Culture| 211 p.
Subject:
62 RADIOLOGY AND NUCLEAR MEDICINE; AGAR; BLOOD; BONE MARROW; BONE MARROW CELLS; COLONY FORMATION; GUINEA PIGS; HAMSTERS; IN VITRO; LIVER; MACROPHAGES; MICE; RABBITS; RADIOSTERILIZATION; RATS; SPLEEN; THERAPEUTIC USES; THERAPY; URINE
OSTI ID:
22192464
Research Organizations:
International Atomic Energy Agency, Vienna (Austria)
Country of Origin:
IAEA
Language:
English
Other Identifying Numbers:
TRN: XA14M0701016953
Submitting Site:
INIS
Size:
page(s) 3-12
Announcement Date:
Feb 20, 2014

Citation Formats

Metcalf, D. Agar Technique for the Cultivation In Vitro of Bone-Marrow Colonies. IAEA: N. p., 1969. Web.
Metcalf, D. Agar Technique for the Cultivation In Vitro of Bone-Marrow Colonies. IAEA.
Metcalf, D. 1969. "Agar Technique for the Cultivation In Vitro of Bone-Marrow Colonies." IAEA.
@misc{etde_22192464,
title = {Agar Technique for the Cultivation In Vitro of Bone-Marrow Colonies}
author = {Metcalf, D.}
abstractNote = {In solid-state agar cultures certain haemopoietic cells proliferate and form discrete colonies of 200 - 4000 cells. Colony formation is dependent on stimulation by the colony-stimulating factor, and this is achieved by (1) the use of a cell feeder layer, (2) the addition of conditioned medium, or (3) the addition of human or mouse serum or urine containing the factor. All colonies initially contain granulocytic cells which differentiate from myeloblasts to polymorphs as colony growth proceeds. Later colonies develop a second population of phagocytic mononuclear cells (macrophages). The colony-forming-system is simple, readily quantitated and highly reproducible. Linear dose responses occur between the dose of colony-stimulating factor and the number and size of colonies developing from a standard number of bone-marrow cells. In-vitro colony formation has been achieved with haemopoietic cells of the following species: mouse, rat, hamster, guinea pig, rabbit and human. In the adult mouse, colony-forming cells are located in the bone marrow, spleen and blood and in the embryo, in the yolk sac, liver and spleen. The colony-forming cell appears to be an early member of the granulocytic series. The colony-forming system has been used as a quantitative assay system: (1) to assay levels of colony-stimulating factor in serum and urine and in the chemical- characterization and purification of the factor; and (2) to enumerate the number of colony-forming cells in haemopoietic tissues in response to a variety of experimental procedures and disease states. Since the system is applicable to human bone-marrow cells, it should prove of value in the quantitative assay of (1) survival of human bone marrow on storage, and (2) bone-marrow content of granulocytic precursor cells in various disease states and following various types of therapy. The system is not suitable for the mass production in vitro of haemopoietic cells for therapeutic use. (author)}
place = {IAEA}
year = {1969}
month = {Jul}
}