Primary culture of mantle cells of the pearl oyster Pinctada mazatlanica (Bivalvia: Pteriidae), with possible application to pearl farming

Cultivo celular de manto de Pinctada mazatlanica

Abstract

Background. The technique for inducing pearl production in mollusks, which relies on the grafting of a round nucleus and a piece of mantle tissue from a donor within the gonad of a recipient, generates high mortality and  rejection  rates.  Goals.  To  develop  methodological  basis  for  in  vitro  primary  cultivation  of  mantle  cells  in the pearl oyster Pinctada mazatlanica, with potential application to pearl production. Methods. Five me-thods for enzymatic dissociation, three different culture media, two cell substrates, and four osmolarities of the culture medium were evaluated. In vitro intake of calcium by mantle tissue was also quantified through histochemistry  and  atomic  absorption  spectrophotometry.  Results.  The  addition  of  collagenase  I  (414  U/mg), at 37°C increased viability and performance of disaggregated cells, which grew better in RPMI-1640 cell medium with poly-D-lysine. Under these conditions, the cells changed from the round-type cell to the fibroblast-type cell at third day and were maintained until day 25. Culture of explants resulted better with RPMI-1640  medium,  at  37  °C,  without  poly-D-lysine,  and  at  0.6  M  NaCl,  where  calcium  absorption  was  evident. Conclusions. These results set the basis of future investigations reformulating new culturing media, evaluating growth factors, and identifying molecules and genes related to biomineralization and synthesis of CaCO3, calcite and aragonite.