Differentiation of human bone marrow‐derived fibroblastoid colony forming cells (CFU‐F) and their roles in haemopoiesis in vitro

Abstract

Human bone marrow-derived fibroblastoid colonies have been quantitatively developed in a liquid culture. A linear relationship between cell number plated and colony number formed supports their clonal origin and hydroxyurea killing indicates that the fibroblastoid colony forming cell (CFU-F) is not in cell cycle in normal bone marrow. Adipose cells were induced in the fibroblastoid colonies by the addition of hydrocortisone (optimal concentration: 10(-6) M). Furthermore, adherent layers with adipocytes provided a more favourable condition for maintaining haemopoiesis in Dexter-system cultures. These results indicate that CFU-F belongs to stromal precursor cells intimately involved in the formation of the haemopoietic microenvironment. Colony incidence of CFU-F was almost normal in most patients with aplastic anaemia, haemopoietic dysplasia and chronic myelogenous leukaemia. However, in acute myelogenous leukaemia, it varied with the stage of the disease. It is concluded that the colony assay is useful for investigating stroma/haemopoietic cell interactions.

Hierarchy of mesenchymal stem cells: Comparison of multipotentmesenchymal stromal cells with fibroblast colony forming units

“…Concentration of CFU-F in bone marrow of healthy donors declined with donors' age (R = −0.68, p = 0.004), that agree with data of other investigators [17][18][19]. The whole group of donors was divided on the basis of age: group younger than median (32 years) was designated as "young", and older than median as "old".…”

Stromal abnormalities in neoplastic bone marrow diseases

“…CML fibroblasts exhibited the same proliferative behavior and protein synthesizing capacity as normal fibroblasts [34]. In the majority of cases reported, the numbers of CFU-F were within the normal range [23,101,114,162,219]. The number of adipocyte colony-forming cells was often increased [101], a finding which is difficult to reconcile with the already mentioned reduction in fat cells in histological sections [130].…”

“…Initially decreased CFU-F numbers tend to normalize upon attainment of complete remission and go back to subnormal levels at relapse (Fig. 2) [114,117,162]. Decreased fibroblast precursor numbers are not necessarily incompatible with increased numbers of mature fibroblasts, as observed in trephine biopsies from AML patients [50], because the stroma system has a very low cell turnover [73,88,89] and the mere enumeration of CFU-F says little about the individual precursor cell's proliferative capacity [222].…”

“…In AML, CFU-F numbers vary widely, with significantly reduced numbers in 30-85% of newly diagnosed cases [99,114,117,154,162,167,219]. A normal CFU-F frequency at diagnosis is associated with a favorable prognosis [99,117].…”

Pulsed electromagnetic fields decrease proinflammatory cytokine secretion (IL-1β and TNF-α) on human fibroblast-like cell culture

“…Using a sterile Pasteur pipette (Labolan, Navarra, Spain), the plasma layer on top was removed and discarded, care being taken to prevent any disturbance to the mononuclear cell layer. Dishes were prepared with 1 9 10 6 MNC in 3 ml of McCoy's 5 A liquid culture medium (AppliChem, Darmstadt, Germany) supplemented with antibiotics (100 UI/ml of penicillin) and discomplemented sterile foetal bovine serum (Invitrogen, Carlsbad, San Diego, California, USA) at 20%, following a modification of the techniques reported by Gordon D and Kaneko [19]. The cultures were kept in a humidified atmosphere with 5% CO 2 and 95% air at 378 C for 21 days.…”