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The study is aimed at developing appropriate media for the mantle explant culture of abalone Haliotis varia. The effect of different media viz., L-15, Ham's F12, M199 was studied in combination with 10% Fetal Calf Serum (FCS) and 10% tissue extracts of gonad, mantle and whole body of abalone, H. varia to understand cell behaviour, cell yield and cell adherence in mantle explant culture of H. varia. Cultures with L-15 media gave better cell yield and M199 promoted better cell adherence. Addition of mantle extract to all media enhanced the cell yield to a maximum followed by whole body extract. Addition of whole body extract facilitated in cell adherence followed by the addition of mantle extract.

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The aim of the present study was to establish a suitable culture system for tissue explants from the mantle of the green mussel, Perna viridis. The experiments were conducted using healthy, live green mussels in the size range of 75 to 110 g collected from Pulicat Lake, Tamil Nadu. Three different culture media namely M199, Leibovitz L-15 and sterile seawater were used to assess the most suitable medium for growth, proliferation and viability of mantle epithelial cells. The effect of the addition of two supplements viz., 10% foetal calf serum (FCS) and 0.1% yeast extract to the culture media was also evaluated. After carefully isolating the pallial layer from the mantle tissue, 1-2 mm2 size explants were successfully cultured in 12-well plates at 25°C for up to 14 days. Cultures were monitored under light and phase contrast objectives in an inverted microscope. Cell counts were made and cell size was measured for each treatment. Cells were observed to migrate from the periphery of the explant within 24 h after initiation of cultures. The liberated cells were mostly round and were either granulocytes or hyalinocytes. Fibroblast-like cells were also observed. Our results showed that proliferation of epithelial cells from mantle tissue was maximum in seawater medium (7.4x104 cells ml-1), followed by L-15 medium (2.55x104 cells ml-1). Average cell size in seawater medium was 10.72 μm and that in L-15 and M199 media was 8.56 and 6.39 μm, respectively. Adherent cells were also more prominent and higher in number in seawater medium. Supplementation of culture media with 10% FCS and 0.1% yeast extract improved both cell proliferation and cell size in all the three culture media. Four concentrations of 0.1% yeast extract (@ 50 μl, 75 μl, 100 μl, 150 μl ml-1 medium) were tested in the present study and best results were obtained with 100 μl ml-1, with respect to both cell counts and size.

Suja, C.P.; Jayasankar, V.; Srinivasa Raghavan, V., and Bosco, K.M., 2019. Microstructural analysis of in vitro nacre formation on shell bead and titanium by cultured mantle tissue of pearl oyster, Pinctada fucata. In: Jithendran, K.P.; Saraswathy, R.; Balasubramanian, C.P.; Kumaraguru Vasagam, K.P.; Jayasankar, V.; Raghavan, R.; Alavandi, S.Nacre, the inner iridescent layer of molluscan shells is an organic-inorganic composite material. The layered arrangement of platelet-shaped CaCO3 crystals and organic matrix in a brick-and-mortar structure makes it a strong, resilient, and iridescent material. In-vitro nacre formation on shell bead by the mantle tissue of the pearl oyster, Pinctada fucata has already been reported. The current study details a microstructural investigation and comparison of in vitro nacre deposition by the mantle tissue of pearl oyster, P. fucata on shell bead and titanium plates. Nacre platelets were observed by Scanning Electron Microscopy (SEM) and its CaCO3 composition was determined by Energy Dispersive X-Ray Spectroscopy (EDS) analysis on both shell bead and titanium plate. Nacre deposits displayed the typical lamellar brick and mortar" arrangement composed of aragonite platelets which form the "bricks" and organic matrix that forms the "mortar". Details of the microstructure of in vitro nacre formed by the mantle tissue of pearl oyster were elucidated for the first time by Atomic Force Microscopy (AFM). Further, the deposition of nacre on titanium proves its potential application as a suitable biomaterial in dental and bone implants, due to its biocompatibility towards regeneration. SEM, AFM, in vitro nacre, microstructure, titanium, biomaterial. ADDITIONAL INDEX WORDS: