title: In-vitro culture system for mesenchymal progenitor cells derived from waste human ovarian follicular fluid.
abstract: To characterize different cell populations in the human ovary, morphological and functional characteristics of cell populations collected during routine IVF procedures were studied. Cells obtained from follicular fluid grew in vitro under minimal medium conditions, without growth factor, including leukaemia-inhibiting factor. Morphological analysis revealed a heterogeneous cell population, with cells displaying a fibroblast-like, epithelial-like and also neuron-like features. Morpho-functional characteristics of fibroblast-like cells were similar to mesenchymal stem cells, and, in particular, were positive for mesenchymal stemness markers, including CD90, CD44, CD105, CD73, but negative for epithelial proteins, such as cytokeratins, CD34 and CD45 antigens. Cell proliferation activity at different times and colony-forming unit capability were evaluated, and multipotency of a subset of granulosa cells was established by in-vitro differentiation studies (e.g. osteogenic, chondrogenic and adipogenic differentiation). This study suggests that cells provided by mesenchymal plasticity can be easily isolated by waste follicular fluid, avoiding scraping of human ovaries, and cultivated in minimal conditions. Successful growth of such progenitor cells on three-dimensional cryogel scaffold provides the basis for future developments in tissue engineering. This culture system may be regarded as an experimental model in which biological behaviour is not influenced by specific growth factors.
authors.affiliation: Department of Public Health, Experimental Medicine and Forensic, Histology and Embryology Unit, University of Pavia, via Forlanini, 10, I-27100 Pavia, Italy;. Electronic address: federica.riva01@unipv.it.
authors.forename: Federica
authors.initials: F
authors.lastname: Riva
authors.affiliation: Research Center for Reproductive Medicine, Obstetrics and Gynecology Unit, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy;
authors.forename: Claudia
authors.initials: C
authors.lastname: Omes
authors.affiliation: Research Center for Reproductive Medicine, Obstetrics and Gynecology Unit, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy;
authors.forename: Roberto
authors.initials: R
authors.lastname: Bassani
authors.affiliation: Research Center for Reproductive Medicine, Obstetrics and Gynecology Unit, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy;; Department of Clinical, Surgical, Diagnostic and Pediatric Sciences, University of Pavia, Italy;; Maternal Infant Department, Obstetrics and Gynecology Unit, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy;
authors.forename: Rossella E
authors.initials: RE
authors.lastname: Nappi
authors.affiliation: IGM-CNR, Histochemistry and Cytometry and Department of Biology and Biotechnology, University of Pavia, Italy.
authors.forename: Giuliano
authors.initials: G
authors.lastname: Mazzini
authors.affiliation: Department of Public Health, Experimental Medicine and Forensic, Histology and Embryology Unit, University of Pavia, via Forlanini, 10, I-27100 Pavia, Italy;
authors.forename: Antonia
authors.initials: A
authors.lastname: Icaro Cornaglia
authors.affiliation: Department of Public Health, Experimental Medicine and Forensic, Histology and Embryology Unit, University of Pavia, via Forlanini, 10, I-27100 Pavia, Italy;
authors.forename: Andrea
authors.initials: A
authors.lastname: Casasco
mesh_terms: D000328:Adult; D015703:Antigens, CD / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D005260:Female; D005307:Fertilization in Vitro; D015571:Follicular Fluid / Q000166:cytology*; D006801:Humans; D007247:Infertility, Female / Q000378:metabolism / Q000473:pathology; D007558:Italy; D008506:Medical Waste; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000473:pathology; D054315:Oocyte Retrieval; D010062:Ovulation Induction; D054457:Tissue Scaffolds
chemical_list: D015703:Antigens, CD; D008506:Medical Waste
keywords: 3D scaffold; granulosa cells; human follicular fluid; in-vitro differentiation; leukemia-inhibiting factor; mesenchymal stem cells