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20171
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Subject: Animal Biotechnology × Author: DHAR, PRIYA × Thesis Type: M.V.Sc. ×
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    ThesisItemOpen Access
    CHARACTERIZATION AND QUALITY EVALUATION OF PIG FIBROBLAST CELLS AT DIFFERENT SUBCULTURES
    (Assam Agricultural University, Khanapara,Guwahati, 2017-07) DHAR, PRIYA; Nath, N. Ch.
    Fibroblast cells are the most common cells of connective tissue. In the present study, porcine fibroblast cells were isolated, cultured and subcultured upto six passages from adult and fetal skin samples. The time required to attain 70% confluence in primary culture of fetal fibroblast cells was found to be 26.40±2.40 hours, which was significantly lower (p≤0.01) than 67±4.80 hours of adult fibroblasts culture. The cells were maintained upto six passages which was 38.8 days for adult and 26 days for fetal fibroblast cells. The fetal fibroblast cells (days) required significantly lower (p≤0.01) time to subculture than adult fibroblast cells. Morphologically the isolated cells had fibroblastic character like typical fusiform shape, turgor vitalis cytoplasm, centrally located nuclei and flame like migration pattern upto sixth passage. The subculture time and cell viability (%) was significantly (p≤0.01) better from third subculture onwards upto sixth subculture as compare to primary and secondary cultures. TUNEL assay revealed a decreasing trend of apoptotic index from primary culture onwards to sixth subculture for both adult and fetal fibroblast cells. Fetal fibroblast cells maintained a lower apoptotic index than adult fibroblast in each of the subcultures. Therefore, fibroblast cells were found to be better from third subculture onwards to sixth subculture in regards to subculture time, viability (%), apoptotic index and in maintaining its normal morphological characteristics. ALP is considered as important marker for identification of pluripotent embryonic stem cells as well as multipotent mesenchymal stem cells. Isolated fibroblast cells of all the subcultures showed activity for ALP test indicating their progressive and undifferentiated quality and may be of multipotent in nature.