Isolamento di cellule stromali mesenchimali da tessuto adiposo di tartaruga palustre americana (Trachemys scripta) prelevato durante intervento di orchiectomia ed ovariosalpingectomia laparoscopica

Mesenchymal stromal cells (MSCs) are of strong clinical interest in veterinary regenerative medicine. Currently, there are no published studies about their isolation and characterization in reptiles. The aim of this work was to evaluate the feasibility of the isolation of adipose tissue-MSCs obtained from male and female pond sliders (Trachemis scripta) during a routine neutering. Five fat tissue samples (obtained from five animals) were collected during elective neutering. Fat fragments were processed, and the cell suspension was harvested and incubated at 28°C in a professional humidified incubator. Cell growth rate was characterized by direct cell counting. The cells were successfully differentiated into adipogenic, chondrogenic and osteogenic lineages. Cell phenotype was characterized by RT-PCR and amplicon sequencing of a panel of markers routinely used for mammalian MSCs characterization. Blood cells were used to validate primers for genes not expressed in MSCs. CFU forming ability was evaluated in low density cell cultures. Cells exhibited the capacity to undergo differentiation into adipocytic, chondrogenic, and osteogenic lineages. The cells exhibited the expression of CD105, CD73, CD44, and CD90, while lacking the expression of CD34 and HLADRA. The sequence homology exceeded 98% with sequences reported for the Trachemys scripta genome. This is the first study regarding the isolation, in vitro expansion and characterization of reptile MSCs obtained from fat tissue. These preliminary data strongly suggest that the isolation of MSCs-like cells from chelonian fat tissue is feasible, opening new prospectives for their potential application in the field of regenerative medicine, for both pet and wild endangered reptile species.