Proteins from cultivated mammary epithelial cells derived from bovine milk

TY - ABST

T1 - Proteins from cultivated mammary epithelial cells derived from bovine milk

AU - Che, Jing

AU - Yue, Yuan

AU - Nielsen, Søren Drud-Heydary

AU - Purup, Stig

AU - Poulsen, Nina Aagaard

AU - Larsen, Lotte Bach

PY - 2024/6

Y1 - 2024/6

N2 - In this study, secretomes from cultivated milk-derived bovine mammary epithelial cells (bMECs) were analyzed to investigate the protein profile of secretions and assess their milk protein synthesis capacity. Briefly, bMECs were isolated from the milk of 5 individual lactating cows and cultivated on Matrigel®-coated inserts in transwell systems, where cells were treated with/without bovine pituitary extract (BPE) in the lower compartment. In a 10-day period, media in lower compartments were collected as lower controls, while components from bMEC were secreted into plain media in upper compartments and collected as secretomes. Proteomics was performed using nLC-timsTOF Pro MS/MS and bioinformatics. More than 900 proteins were identified in the secretomes, where no significant differences were found between the two treatments with/without BPE in terms of the number of proteins identified. This indicates that milk-derived bMECs were capable of secreting similar protein profile without lactogenic stimulation. Moreover, the significantly higher number of identified proteins in secretomes compared to lower controls (283±49 from non-BPE treatment) suggests a separation of upper and lower compartments. Importantly, bMECs showed the secretion of more than 100 milk components, including caseins, whey proteins, enzymes, and other proteins. Despite the identification of interesting proteins, the total protein contents of the secretomes were much lower than regular milk, with caseins taking less than 1% of the total protein abundance. In conclusion, our proteomic study provides insight into the composition of milk-derived bMECs secretomes, indicating this in vitro culture model remains challenged in many aspects, such as upscale and sufficient lactogenesis.

AB - In this study, secretomes from cultivated milk-derived bovine mammary epithelial cells (bMECs) were analyzed to investigate the protein profile of secretions and assess their milk protein synthesis capacity. Briefly, bMECs were isolated from the milk of 5 individual lactating cows and cultivated on Matrigel®-coated inserts in transwell systems, where cells were treated with/without bovine pituitary extract (BPE) in the lower compartment. In a 10-day period, media in lower compartments were collected as lower controls, while components from bMEC were secreted into plain media in upper compartments and collected as secretomes. Proteomics was performed using nLC-timsTOF Pro MS/MS and bioinformatics. More than 900 proteins were identified in the secretomes, where no significant differences were found between the two treatments with/without BPE in terms of the number of proteins identified. This indicates that milk-derived bMECs were capable of secreting similar protein profile without lactogenic stimulation. Moreover, the significantly higher number of identified proteins in secretomes compared to lower controls (283±49 from non-BPE treatment) suggests a separation of upper and lower compartments. Importantly, bMECs showed the secretion of more than 100 milk components, including caseins, whey proteins, enzymes, and other proteins. Despite the identification of interesting proteins, the total protein contents of the secretomes were much lower than regular milk, with caseins taking less than 1% of the total protein abundance. In conclusion, our proteomic study provides insight into the composition of milk-derived bMECs secretomes, indicating this in vitro culture model remains challenged in many aspects, such as upscale and sufficient lactogenesis.

UR - https://www.internationalcellag.com/abstracts

M3 - Conference abstract for conference

T2 - 2nd International Cellular Agriculture Conference, Aarhus, Denmark.

Y2 - 12 June 2024 through 13 July 2024

ER -