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• W2077563008 abstract "Adult wound repair traits including inflammation, fibroplasia, and collagen deposition are not seen at fetal wound sites. This observation raised questions about regulatory mechanisms extent in fetal healing. Transforming growth factor beta (TGF-β) is an important regulatory polypeptide known to orchestrate fibroplasia and collagen synthesis during adult wound repair. Previous studies have suggested that the wounded rabbit fetus is capable of responding with these adult characteristics if provided with exogenous TGF-β. In order to test whether the observed in vivo effects of TGF-β in the rabbit fetus might be due to a direct effect on the fibroblast, TGF-β receptor binding characteristics of early passage cultured embryonic (14 days' gestation), fetal (24 days' gestation), and adult rabbit fibroblasts were studied by flow cytometry. Experiments were carried out using fluorescein-conjugated TGF-β (F-TGF-β) with analysis on an EPICS V flow cytometer. F-TGF-β was incubated with each of the three fibroblast types at 37°C after which time the cells were washed twice and analyzed with a minimum of 105 cells for each data point. F-TGF-β bound rapidly and reversibly to the embryonic, fetal, and adult fibroblasts with saturation being achieved at 1 nmol/L for fetal and adult cells, and 8 nmol/L in the embryonic fibroblasts. Saturating concentrations of F-TGF-β yielded mean channel numbers (a function of relative amounts of F-TGF-β-bound) of 172, 114, and 97 for embryonic, fetal, and adult cells, respectively. The kinetics of receptor binding were rapid, with>80% of the free TGF-β binding sites being occupied in the first two minutes of incubation, except for the adult cells, in which 73% of the free sites were occupied after two minutes. Greater than 95% of the free TGF-β sites were occupied in all three cell types after 25 minutes, and saturation occurred at one hour. These results demonstrate that both embryonic and fetal rabbit fibroblasts possess cell surface receptors for TGF-β previously shown to be present in their adult counterparts. As fibroblasts differentiate during development from embryo through fetus on to adult, there is a progressive reduction in the number of these cell surface receptors. These findings correlate well with recent histologic evidence documenting an abundance of TGF-β in mouse embryo mesenchyma, thus supporting the concept that TGF-β exerts regulatory control during embryogenesis. The current in vitro demonstration of the abundant presence of cell surface receptors for TGF-β in both embryonic and fetal fibroblasts, yet an apparent lack of fibrosis and collagen accumulation following fetal injury, suggests that the availability of TGF-β in vivo is tightly controlled. Adult wound repair traits including inflammation, fibroplasia, and collagen deposition are not seen at fetal wound sites. This observation raised questions about regulatory mechanisms extent in fetal healing. Transforming growth factor beta (TGF-β) is an important regulatory polypeptide known to orchestrate fibroplasia and collagen synthesis during adult wound repair. Previous studies have suggested that the wounded rabbit fetus is capable of responding with these adult characteristics if provided with exogenous TGF-β. In order to test whether the observed in vivo effects of TGF-β in the rabbit fetus might be due to a direct effect on the fibroblast, TGF-β receptor binding characteristics of early passage cultured embryonic (14 days' gestation), fetal (24 days' gestation), and adult rabbit fibroblasts were studied by flow cytometry. Experiments were carried out using fluorescein-conjugated TGF-β (F-TGF-β) with analysis on an EPICS V flow cytometer. F-TGF-β was incubated with each of the three fibroblast types at 37°C after which time the cells were washed twice and analyzed with a minimum of 105 cells for each data point. F-TGF-β bound rapidly and reversibly to the embryonic, fetal, and adult fibroblasts with saturation being achieved at 1 nmol/L for fetal and adult cells, and 8 nmol/L in the embryonic fibroblasts. Saturating concentrations of F-TGF-β yielded mean channel numbers (a function of relative amounts of F-TGF-β-bound) of 172, 114, and 97 for embryonic, fetal, and adult cells, respectively. The kinetics of receptor binding were rapid, with>80% of the free TGF-β binding sites being occupied in the first two minutes of incubation, except for the adult cells, in which 73% of the free sites were occupied after two minutes. Greater than 95% of the free TGF-β sites were occupied in all three cell types after 25 minutes, and saturation occurred at one hour. These results demonstrate that both embryonic and fetal rabbit fibroblasts possess cell surface receptors for TGF-β previously shown to be present in their adult counterparts. As fibroblasts differentiate during development from embryo through fetus on to adult, there is a progressive reduction in the number of these cell surface receptors. These findings correlate well with recent histologic evidence documenting an abundance of TGF-β in mouse embryo mesenchyma, thus supporting the concept that TGF-β exerts regulatory control during embryogenesis. The current in vitro demonstration of the abundant presence of cell surface receptors for TGF-β in both embryonic and fetal fibroblasts, yet an apparent lack of fibrosis and collagen accumulation following fetal injury, suggests that the availability of TGF-β in vivo is tightly controlled." @default.
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• W2077563008 date "1989-08-01" @default.
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• W2077563008 title "Analysis of transforming growth factor beta receptor binding in embryonic, fetal, and adult rabbit fibroblasts" @default.
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