Differentiation of rat neural tissue in a serum-free embryo culture model followedby in vivo transplantation.
Journal Title: Croatian Medical Journal - Year 2002, Vol 42, Issue 6
Abstract
AIM: To analyze neural tissue differentiation in a unique, chemically-definedin vitro culture model of gastrulating rat embryo proper by use of transmission electron microscopy (TEM),proliferating cell nuclear antigen (PCNA) expression, and in vivo transplantation after a 2-week culturein serum-free or serum-supplemented media. Influence of protein-free medium unfavorable for differentiationof neural tissue in vitro was compared with favorable serum-free media enriched with transferrin or albumin.Differentiation of mesodermal derivatives in transplants was also investigated. METHODS: We cultivated9.5-day-old Fischer rat embryos on the gas-liquid interface in the protein-free Eagle's Minimum EssentialMedium (MEM), in MEM with either iron-saturated holotransferrin (50 microg/mL) or iron-free apotransferrin(50 microg/mL), and in medium saturated with either bovine serum albumin (BSA) (4 mg/mL or 400 microg/mL)or rat serum (50%). After the two-week culture period, light microscopy, TEM, and immunohistochemicalmethod for detection of PCNA were done. Some explants were transplanted under the kidney capsule of adultmale rats to be cultured in vivo for additional two weeks. Chi-square test or Fisher exact test wereused to compare the proportion of tissues developed. RESULTS: Proportion of differentiated neural tissuewas similar in explants cultivated in apotransferrin- and holotransferrin-supplemented media (13/33 and9/20, respectively), but higher than in explants cultivated in protein-free medium (1/13). Neurons andglia cells produced a neuropil structure. Myelinization occurred only in serum-supplemented medium. PCNAexpression was detected in a small number of neural tissue cells, even in serum-free cultivated embryos.Differentiation of brain-like tissue, cerebrospinal, and vegetative ganglionic cells occurred in allgroups of transplants. However, in the transplants derived from protein-free medium, the proportion ofneural tissue, cartilage, bone, skeletal and smooth muscle was significantly lower than in transferrin-supplementedmedia (p
Authors and Affiliations
T Belovari, F Bulić-Jakus, G Jurić-Lekić, S Marić, D Jezek, M Vlahović
Keywords
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