title: Use of Stirred Suspension Bioreactors for Male Germ Cell Enrichment.
abstract: Spermatogenesis is a stem cell based system. Both therapeutic and biomedical research applications of spermatogonial stem cells require a large number of cells. However, there are only few germ line stem cells in the testis, contained in the fraction of undifferentiated spermatogonia. The lack of specific markers makes it difficult to isolate these cells. The long term maintenance and proliferation of nonrodent germ cells in culture has so far been met with limited success, partially due to the lack of highly enriched starting populations. Differential plating, which depends on the differential adhesion properties of testicular somatic and germ cells to tissue culture dishes, has been the method of choice for germ cell enrichment, especially for nonrodent germ cells. However, for large animals, this process becomes labor intensive and increases variability due to the need for extensive handling. Here, we describe the use of stirred suspension bioreactors, as a novel system for enriching undifferentiated germ cells from 1-week-old pigs. This method capitalizes on the adherent properties of somatic cells within a controlled environment, thus promoting the enrichment of progenitor cells with minimal handling and variability.
authors.affiliation: Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, Cumming School of Medicine, University of Calgary, 3330 Hospital Drive NW, HMRB 404, Calgary, AB, Canada, T2N 4N1.|Department of Biochemistry and Molecular Biology, Cumming School of Medicine, University of Calgary, 3330 Hospital Drive NW, HMRB 404, Calgary, AB, Canada, T2N 4N1.
authors.forename: Sadman
authors.initials: S
authors.lastname: Sakib
authors.affiliation: Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, Cumming School of Medicine, University of Calgary, 3330 Hospital Drive NW, HMRB 404, Calgary, AB, Canada, T2N 4N1.
authors.forename: Camila
authors.initials: C
authors.lastname: Dores
authors.affiliation: Department of Biochemistry and Molecular Biology, Cumming School of Medicine, University of Calgary, 3330 Hospital Drive NW, HMRB 404, Calgary, AB, Canada, T2N 4N1.|Department of Oncology and Medical Genetics, Cumming School of Medicine, University of Calgary, 3330 Hospital Drive NW, HMRB 404, Calgary, AB, Canada, T2N 4N1.
authors.forename: Derrick
authors.initials: D
authors.lastname: Rancourt
authors.affiliation: Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, Cumming School of Medicine, University of Calgary, 3330 Hospital Drive NW, HMRB 404, Calgary, AB, Canada, T2N 4N1. idobrins@ucalgary.ca.|Department of Biochemistry and Molecular Biology, Cumming School of Medicine, University of Calgary, 3330 Hospital Drive NW, HMRB 404, Calgary, AB, Canada, T2N 4N1. idobrins@ucalgary.ca.
authors.forename: Ina
authors.initials: I
authors.lastname: Dobrinski
mesh_terms: D000818:Animals; D019149:Bioreactors; D002452:Cell Count; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D049109:Cell Proliferation; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D002478:Cells, Cultured; D004867:Equipment Design; D008297:Male; D013091:Spermatogenesis; D013093:Spermatogonia / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology*; D013552:Swine; D013737:Testis / Q000166:cytology
keywords: Germ cells; Spermatogonial stem cell; Spinner flasks; Stirred suspension bioreactor; Testes