ConcePTION WP3 task 3.2. Isolation and characterization of Mammary Epithelial Cells from Göttingen minipig (mpMECs): an in vitro study to compare mpMECs vs pMECs (porcine Mammary Epithelial Cells)

This data set contains data related to the characterization of an appropriate in vitro animal model based on primary culture of Göttingen Minipig Mammary Epithelial Cells (mpMECs). The objective of the research was to verify the accuracy of mpMECs as solid translational model for the study of mammary epithelial barrier and compare mpMECs results with those obtained from Mammary Epithelial Cells isolated from hybrid commercial pig (pMECs). The results showed that it was possible to isolate, culture and expand three pure epithelial cell lines obtained from three different animals. The mpMECs maintained until P10 showed a typical cobblestone morphology and a similar doubling time profile (MG9 32.6 ± 4 h, MG11 30.5 ± 3 h and MG12 27.4 ± 2.8 h, as reported in the data file CONCEPTION_WP3_mpMECs_DoublingTime_09032023.xlsx). DNA index was normal for all the three cell populations with a mean value of 0.98 ± 0.01. Regarding the cell cycle, the cell populations MG9, MG11 and MG12 showed the three distinct phases that could be recognized in a proliferating cell population: G0/G1, S and G2/M (see data file CONCEPTION_WP3_mpMECs_CellCycle_DNAIndex_09032023.xlsx). All the three mpMECs cell lines expressed epithelial markers Epithelial-Cadherin (E-Cad) and Cytokeratin 18 (CK18), confirming their epithelial origin (see data file CONCEPTION_WP3_mpMECs_FlowCytometer_E-Cad_CK18_09032023.xlsx). In particular in MG12, the contour of CK18 positive peak showed a shoulder suggesting the presence of a cellular subpopulation with a particularly high positivity. The barrier function of mpMECs was evaluated via TEER and fluorescein sodium (SF) transport, the formation of the monolayer integrity was evaluated in all the three primary mpMECs and in the pre-mixed pool of mpMECs and pMECs. MG9, MG11 and MG12 resulted in a similar TEER profile at the 0.15×10^6 cells, achieving higher TEER and lower SF values at the day 3 and 4 of culture, which was also confirmed in the pre-mixed pool case. Moreov