Immortalized mouse caput epididymal epithelial (mECap18) cell line recapitulates the in-vivo environment.
Jess E MulhallNatalie A TriggIlana R BernsteinAmanda L AndersonHeather C MurrayPetra SipiläTessa LordJohn E SchjenkenBrett NixonDavid A Skerrett-ByrnePublished in: Proteomics (2023)
Residing between the testes and the vas deferens, the epididymis is a highly convoluted tubule whose unique luminal microenvironment is crucial for the functional maturation of spermatozoa. This microenvironment is created by the combined secretory and resorptive activity of the lining epididymal epithelium, including the release of extracellular vesicles (epididymosomes), which encapsulate fertility modulating proteins and a myriad of small non-coding RNAs (sncRNAs) that are destined for delivery to recipient sperm cells. To enable investigation of this intercellular communication nexus, we have previously developed an immortalized mouse caput epididymal epithelial cell line (mECap18). Here, we describe the application of label-free mass spectrometry to characterize the mECap18 cell proteome and compare this to the proteome of native mouse caput epididymal epithelial cells. We report the identification of 5,313 mECap18 proteins, as many as 75.8% of which were also identified in caput epithelial cells wherein they mapped to broadly similar protein classification groupings. Furthermore, key pathways associated with protein synthesis (e.g., EIF2 signaling) and cellular protection in the male reproductive tract (e.g., sirtuin signaling) were enriched in both proteomes. This comparison supports the utility of the mECap18 cell line as a tractable in-vitro model for studying caput epididymal epithelial cell function.
Keyphrases
- label free
- mass spectrometry
- stem cells
- induced apoptosis
- single cell
- machine learning
- deep learning
- signaling pathway
- liquid chromatography
- cell cycle arrest
- cell death
- oxidative stress
- bone marrow
- mesenchymal stem cells
- capillary electrophoresis
- small molecule
- amino acid
- protein protein
- cell proliferation
- high performance liquid chromatography
- tandem mass spectrometry