engleski

Modified embryo organ culture model system for preclinical studies of growth factor-induced neural tissue differentiation in mammals

Aims: Fibroblast growth factor (FGF) and nerve growth factor (NGF) are found in various parts of human and vertebrate embryos during development. The development of the vertebrate nervous system begins with the inductive differentiation of the neural plate from the dorsal side of flat ectoderm at the end of gastrulation. The role of FGF and NGF in critical period for mammalian development, gastrulation, was studied. Material and methods: In our laboratory a unique organ culture system of mammalian embryo was designed. Embryonic parts of the whole gastrulating rat embryos were cultivated on the stainless-steel grid supported lens paper, at the air-liquid interface during two weeks using liquid serum-free and protein-free medium. A signal-protein FGF or NGF (100 ng/ml and 200 ng/ml) were added, or both together in culture medium in time frame of 2, 5 or 9 days. Results: Embryos developed into teratoma built of intermixed tissues, including nerve tissue. In control serum-free medium neuroblasts were absent. NGF did never improve differentiation of neuroblasts. FGF significantly stimulated the differentiation of neural tissue during 5 days, and the best during 9 days. In FGF/NGF-treated embryos differentiation of neuroblasts was the same as was in FGF-treated. Conclusions: Our model system is simpler than the embryo in vivo, but closer to normal development than the cell cultures used in other studies. The neuroblasts found were direct answer to molecular signals added. This fact is in accordance with the idea that in a multi-level process of neural differentiation, FGF is the neural inducer in mammals.

embryo culture; growth factor; differentiation

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