Marrow Reticulo-Fibroblastoid Colonies (CFU-RF Derived) Spontaneously Release an Erythroid Colony (BFU-E) Enhancing Factor

  • Carlos A. Izaguirre
  • William M. Ross
  • Elizabeth Y. Hsu
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 34)


The marrow microenvironment is composed of an extracellular matrix as well as a heterogeneous population of cells. Isolation of the various cell types and analysis of their function is necessary for a better understanding of their roles in hemopoiesis. We have recently reported a colony assay for a cellular component of the marrow microenvironment. The assay consists of a plasma clot-methylcellulose marrow culture. The stimulator is PHA-stimulated leukocyte conditioned medium (PHA-LCM) and hydrocortisone (5 x 10-5M). The fibrin strands appear to act as a substrate for the growth of Reticulo-Fibroblastoid colonies derived from the CFU-RF precursor. RF colonies can be subcultured forming adherent layers when transferred to liquid cultures. Confluent adherent layers can be maintained for long periods of time by changing medium every 3 to 5 days. Supernatants derived from unstimulated RF cultures (RF-CM) were tested for growth promotion of hemopoietic precursors. We found: (1) RF-CM by itself does not induce colony formation. (2) In the presence of erythropoietin, RF-CM enhances the growth of BFU-E. (3) Recombinant IL 4 also enhances BFU-E formation, but in our assays IL 4 induced fewer colonies than RF-CM and the colonies were smaller. (4) Because neither IL 4 nor RF-CM, by themselves, can stimulate colony formation, we compared the effect of RF-CM on assays that are known to show other IL 4 functions. RF-CM did not induce proliferation of PHA induced blast T cells, a known property of IL 4. In conclusion, RF cells in culture appear to secrete, spontaneously, a factor that enhances BFU-E formation, and, unlike GM-CSF and 1L3, RF-CM has no effect on other hemopoietic colony forming cells. IL 4 has similar properties, but RF-CM appears not to have the B- and T-cell growth factor activities of Interleukin 4.


Colony Formation Bone Marrow Culture Hemopoietic Stem Cell Adherent Layer Human Bone Marrow Cell 


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Copyright information

© Plenum Press, New York 1988

Authors and Affiliations

  • Carlos A. Izaguirre
    • 1
  • William M. Ross
    • 2
    • 4
  • Elizabeth Y. Hsu
    • 3
  1. 1.Department of MedicineUniversity of OttawaOttawaCanada
  2. 2.Department of PhysiologyUniversity of OttawaOttawaCanada
  3. 3.Department of PediatricsUniversity of OttawaOttawaCanada
  4. 4.Defense Research EstablishmentOttawaCanada

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