| Summary: | 碩士 === 國立台北師範學院 === 數理教育研究所 === 89 === TB cell line derived from brain tissue of a Tilapia was established in L-15 medium containing 10﹪fetal bovine serum. The cell line was characterized by morphological observation, growth curve, chromosome number distribution, the infection of virus and immunostaining. Morphology of the TB cells presented as epithelial-like cells, fibroblast-like cells and neuron-like cells. Growth curve experiment revealed four phases as lag phase, logarithmic growth phase, stationary phase, death phase. Result of chromosome analysis revealed chromosome number of the TB cell line distributed from 21 to 44, with a mode number of 26, 28, 39. TB cells have persistent HCRV(hard clam reovirus) infection. HCRV was able to replicate in TB cell line with clear CPE(cytopathogenic effect), as evidenced by the presence of about 106.1 TCID50/0.1ml virus in the cell culture supernatants. Results of immunostaining showed keratin immunoreactivity significantly and their expression is typical of epithelial differentiation. Immunostaining of NF200 and Tublin-β, neuron markers, showed positive in TB cell line. TH and DDC typical of dopamine neurons were also positive.
Neuronal differentiation of TB cells was shown to occur spontaneously after treated with All Trans Retinoic Acid(ATRA, 10μM), in the absence of any added cytokines or special growth factors. Results of TEM observation have shown that many neurofilaments increased in the cytoplasm. Results of SEM observation, the cell bodies were small and rough with several long neuritis to form a complex network structure. Thus, ATRA has influence on the TB cells from the CNS, and make the morphorlogy of the TB cells be more mature.
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