title: Epithelialization and stromalization of porcine follicular granulosa cells during real-time proliferation - a primary cell culture approach.
abstract: The process of oocyte growth and development takes place during long stages of folliculogenesis and oogenesis. This is accompanied by biochemical and morphological changes, occurring from the preantral to antral stages during ovarian follicle differentiation. It is well known that the process of follicle growth is associated with morphological modifications of theca (TCs) and granulosa cells (GCs). However, the relationship between proliferation and/or differentiation of porcine GCs during long-term in vitro culture requires further investigation. Moreover, the expression of cytokeratins and vimentin in porcine GCs, in relation to real-time cell proliferation, has yet to be explored. Utilizing confocal microscopy, we analyzed cytokeratin 18 (CK18), cytokeratin 8 + 18 + 19 (panCK), and vimentin (Vim) expression, as well as their protein distribution, within GCs isolated from slaughtered ovarian follicles. The cells were cultured for 168 h with protein expression and cell proliferation index analyzed at 24-h intervals. We found the highest expression of CK18, panCK, and Vim occurred at 120 h of in vitro culture (IVC) as compared with other experimental time intervals. All of the investigated proteins displayed cytoplasmic distribution. Analysis of real-time cell proliferation revealed an increased cell index after the first 24 h of IVC. Additionally, during each period between 24-168 h of IVC, a significant difference in the proliferation profile, expressed as the cell index, was also observed. We concluded that higher expression of vimentin at 120 h of in vitro proliferation might explain the culmination of the stromalization process associated with growth and domination of stromal cells in GC culture. Cytokeratin expression within GC cytoplasm confirms the presence of epithelial cells as well as epithelial-related GC development during IVC. Moreover, expression of both cytokeratins and vimentin during short-term culture suggests that the process of GC proliferation is also highly associated with porcine ovarian follicular granulosa cell differentiation in vitro.
authors.affiliation: Department of Histology and Embryology, Poznan University of Medical Sciences, Poznan, Poland.
authors.forename: S
authors.initials: S
authors.lastname: Ciesiółka
authors.affiliation: Department of Anatomy, Poznan University of Medical Sciences, Poznan, Poland.
authors.forename: A
authors.initials: A
authors.lastname: Bryja
authors.affiliation: Department of Histology and Embryology, Poznan University of Medical Sciences, Poznan, Poland.
authors.forename: J
authors.initials: J
authors.lastname: Budna
authors.affiliation: Department of Anatomy, Poznan University of Medical Sciences, Poznan, Poland.
authors.forename: W
authors.initials: W
authors.lastname: Kranc
authors.affiliation: Department of Histology and Embryology, Poznan University of Medical Sciences, Poznan, Poland.
authors.forename: A
authors.initials: A
authors.lastname: Chachuła
authors.affiliation: Institute of Veterinary Sciences, Poznan University of Life Sciences, Poznan, Poland.
authors.forename: D
authors.initials: D
authors.lastname: Bukowska
authors.affiliation: Department of Toxicology, Poznan University of Medical Sciences, Poznan, Poland.
authors.forename: H
authors.initials: H
authors.lastname: Piotrowska
authors.affiliation: Department of Anatomy, Poznan University of Medical Sciences, Poznan, Poland.
authors.forename: L
authors.initials: L
authors.lastname: Porowski
authors.affiliation: Institute of Veterinary Sciences, Poznan University of Life Sciences, Poznan, Poland.
authors.forename: P
authors.initials: P
authors.lastname: Antosik
authors.affiliation: Department of Anatomy, Poznan University of Medical Sciences, Poznan, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Bruska
authors.affiliation: Institute of Veterinary Sciences, Poznan University of Life Sciences, Poznan, Poland.
authors.forename: K P
authors.initials: KP
authors.lastname: Brüssow
authors.affiliation: Department of Histology and Embryology, Poznan University of Medical Sciences, Poznan, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Nowicki
authors.affiliation: Department of Histology and Embryology, Poznan University of Medical Sciences, Poznan, Poland; Department of Histology and Embryology, Wroclaw Medical University, Wroclaw, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Zabel
authors.affiliation: Department of Histology and Embryology, Poznan University of Medical Sciences, Poznan, Poland; Department of Anatomy, Poznan University of Medical Sciences, Poznan, Poland.
authors.forename: B
authors.initials: B
authors.lastname: Kempisty
mesh_terms: D000818:Animals; D002455:Cell Division; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology; D005260:Female; D006107:Granulosa Cells / Q000166:cytology* / Q000378:metabolism; D007633:Keratins / Q000096:biosynthesis / Q000235:genetics; D018613:Microscopy, Confocal; D009866:Oogenesis; D006080:Ovarian Follicle / Q000166:cytology; D061251:Primary Cell Culture; D017154:Stromal Cells / Q000166:cytology; D034421:Sus scrofa; D013552:Swine; D014746:Vimentin / Q000096:biosynthesis / Q000235:genetics
chemical_list: D014746:Vimentin; D007633:Keratins