- Email: [email protected]
Biochemical, and biological characterization of bFGF extracted from human placenta
lbstrarts:
European Placenta Group
-497
ma1 implantation sites appears to support the general concept that premenstrual conception is a frequent phenomenon in the background of implantations outside the usual sites of nidation, thus supporting the basic premise of the ‘post-mid-cycle theory’ concerning etiology and pathological mechanism.
CRUDE PLACENTAL EXTRACT WITH GROWTH FACTOR ACTIVITY (EAP): BIOCHEMICAL AND BIOLOGICAL CHARACTERIZATION J. Tiollier, S. Uhlrich, M. Tardy &J. L. Tayot (IMEDEX, B.P. 30, Z.I. des Troques, 69630 Chaponost, France) A crude extract (EAP) is prepared at an industrial scale from human placental tissue. The protein content of this extract was characterized by polyacrylamide gel electrophoresis and immunodiffusion techniques. Its content in growth factors, such as EGF, IGFs, BTGF, PDGF and FGFs, was estimated by specific techniques designed for each growth factor. The biological characterization of growth factor activity of EAP was performed by cell culture assay. EAP stimulates the proliferation of various cells (MRC5 and various cutaneous fibroblasts, cornea1 endothelial cells). The ED,, for stimulation of proliferation by fibroblasts was found to be in the range of loo-zoopg/ml. The main biological properties of EAP, observed in vitro, include replacement of serum in cell culture media, stimulation of cell migration (endothelial cells and keratinocytes). In addition, the angiogenic activity of EAP is demonstrated in chick chorioallantoic membrane.
AND BIOLOGICAL CHARACTERIZATION OF bFGF BIOCHEMICAI, EXTRACTED FROM HUMAN PLACENTA S. Uhlrich, J. Tiollier, M. Tardy &J. L. Tayot (IMEDEX, B.P. 38 - Z.I. Des Troques, 69630 Chaponost, France) Basic Fibroblast Growth Factor is a multifunctional polypeptide which controls the proliferation and differentiation of mesoderm- and neuroectoderm-cells. We report here the quantitative purification of bFGF from placental tissue. The bFGF fraction was purified to homogeneity in two chromatographic steps. The yield of purification was IO pg bFGF per kg of placenta. This fraction consists of two proteins with apparent molecular weights 16 ooo and 18 ooo. The two molecular forms were separated by gel electrophoresis followed by reverse-phase HPLC. Both proteins have the same N-terminal sequence: Pro-Ala-Leu-Pro-Glu-Asp-Gly-Gly-Ser-Gly-Ama-Phe . . which is identical to that ofhuman bFGF. The nature of the molecular difference between these two forms of bFGF will be discussed. The biological characterization of growth factor activity of these molecules was performed by cell culture assay. The ED,, for stimulation of proliferation of CCL39 fibroblasts or bovine cornea1 endothelial cells has been found to be in the range of 0.5-1 ng/ml for basic FGF. Cell growth is dependent on the concentration of FGF present in the medium. The main properties of bFGF, obseived in vitro, include stimulation of cell migration, acceleration of cellular proliferation in serum-free media and prolongation of life span of primary cultures of bovine cornea1 endothelial cells.
European Placenta Group
-497
ma1 implantation sites appears to support the general concept that premenstrual conception is a frequent phenomenon in the background of implantations outside the usual sites of nidation, thus supporting the basic premise of the ‘post-mid-cycle theory’ concerning etiology and pathological mechanism.
CRUDE PLACENTAL EXTRACT WITH GROWTH FACTOR ACTIVITY (EAP): BIOCHEMICAL AND BIOLOGICAL CHARACTERIZATION J. Tiollier, S. Uhlrich, M. Tardy &J. L. Tayot (IMEDEX, B.P. 30, Z.I. des Troques, 69630 Chaponost, France) A crude extract (EAP) is prepared at an industrial scale from human placental tissue. The protein content of this extract was characterized by polyacrylamide gel electrophoresis and immunodiffusion techniques. Its content in growth factors, such as EGF, IGFs, BTGF, PDGF and FGFs, was estimated by specific techniques designed for each growth factor. The biological characterization of growth factor activity of EAP was performed by cell culture assay. EAP stimulates the proliferation of various cells (MRC5 and various cutaneous fibroblasts, cornea1 endothelial cells). The ED,, for stimulation of proliferation by fibroblasts was found to be in the range of loo-zoopg/ml. The main biological properties of EAP, observed in vitro, include replacement of serum in cell culture media, stimulation of cell migration (endothelial cells and keratinocytes). In addition, the angiogenic activity of EAP is demonstrated in chick chorioallantoic membrane.
AND BIOLOGICAL CHARACTERIZATION OF bFGF BIOCHEMICAI, EXTRACTED FROM HUMAN PLACENTA S. Uhlrich, J. Tiollier, M. Tardy &J. L. Tayot (IMEDEX, B.P. 38 - Z.I. Des Troques, 69630 Chaponost, France) Basic Fibroblast Growth Factor is a multifunctional polypeptide which controls the proliferation and differentiation of mesoderm- and neuroectoderm-cells. We report here the quantitative purification of bFGF from placental tissue. The bFGF fraction was purified to homogeneity in two chromatographic steps. The yield of purification was IO pg bFGF per kg of placenta. This fraction consists of two proteins with apparent molecular weights 16 ooo and 18 ooo. The two molecular forms were separated by gel electrophoresis followed by reverse-phase HPLC. Both proteins have the same N-terminal sequence: Pro-Ala-Leu-Pro-Glu-Asp-Gly-Gly-Ser-Gly-Ama-Phe . . which is identical to that ofhuman bFGF. The nature of the molecular difference between these two forms of bFGF will be discussed. The biological characterization of growth factor activity of these molecules was performed by cell culture assay. The ED,, for stimulation of proliferation of CCL39 fibroblasts or bovine cornea1 endothelial cells has been found to be in the range of 0.5-1 ng/ml for basic FGF. Cell growth is dependent on the concentration of FGF present in the medium. The main properties of bFGF, obseived in vitro, include stimulation of cell migration, acceleration of cellular proliferation in serum-free media and prolongation of life span of primary cultures of bovine cornea1 endothelial cells.