Area 3: Molecular genetics of eukaryotes Biological characterization and in vitro culture of human spermatogonial stem cells Elena Vicini Associate Professor Department of Anatomical, Histological, Forensic & Orthopaedic Sciences Tel: +39 06 4976 /6804 e‐mail: [email protected] ABSTRACT In the male gonads of mammals, the germline stem cells belong to a subpopulation of undifferentiated spermatogonia localized in the basal layer of the seminiferous epithelium. While, spermatogenesis in rodents and primates, including humans, has been considered dissimilar with important biological differences in the identity and behavior of stem cells, recent studies from our laboratory and others recently challenged this concept. Expect for the existence of type Apale and Adark spermatogonia, and that Adark spermatogonia are quiescient not much is known about the spermatogonial compartment in humans. Clearly, there is need of new data in order to obtain insight in the regulation of human germline compartment. In this direction, a new re‐assesment of the stages of the seminiferous epithelium (cellular associations), the analysis of spermatogonial behaviors in whole mounted seminiferous tubules, as well as development of spermatogonial stem cells lines are highly warranted. Feasibility of this proposal strongly relay on the availability of unfixed human testicular cells. In this direction, we have already obtained human testicular cells taken from healthy heart beating organ donors. Data obtained will be highly relevant to the study of molecular mechanisms regulating early step of spermatogenesis, for future applications on male infertility, and to unveil molecular mechanisms underlying pluripotency. GROUP COMPONENTS ‐ Carla Boitani, associate professor, Sapienza University of Rome ‐ Barbara Muciaccia, postdoctoral fellow, Sapienza University of Rome ‐ Mario Stefanini, senior professor, Sapienza University of Rome 1
‐ Anna Maria Lustri, Ph.D. student, Sapienza University of Rome ‐ Dante Lamberti, Ph.D. student, Sapienza University of Rome ‐ Stefania Fera, laboratory technician, Sapienza University of Rome RELEVANT PUBBLICATIONS (2008‐2012) ‐ Grisanti L, Corallini S, Fera S, Muciaccia B, Stefanini M, Witke W, Vicini E (2009). Inactivation of Numb and Numblike in spermatogonial stem cells by cell‐permeant Cre recombinase. Differentiation, 78: 131‐136. IF 3.06 ‐ Grisanti L, Falciatori I, Grasso M, Dovere L, Fera S, Muciaccia B, Fuso A, Berno V, Boitani C, Stefanini M, Vicini E. (2009). Identification of Spermatogonial Stem Cell Subsets by Morphological Analysis and Prospective Isolation. Stem Cells, 27: 3043‐3052. IF 7.87 ‐ Scaldaferri ML, Fera S, Grisanti L, Sanchez M, Stefanini M, De Felici M, Vicini E (2011). Identification of side population cells in mouse primordial germ cells and prenatal testis. The international Journal of Developmental Biology 55: 209‐14. IF 2.85 ‐ Grasso M., Fuso A., Dovere L.,. de Rooij D., Stefanini M., Boitani C. and Vicini E. (2012). Distribution of GFRA1‐expressing spermatogonia in adult mouse testis. Reproduction 143(3):325‐32. IF 3.05 ‐ Ferranti F, Muciaccia B, Ricci G, Dovere L, Canipari R, Magliocca F, Stefanini M, Catizone A, Vicini E. (2012) Glial cell line‐derived neurotrophic factor promotes invasive behaviour in testicular seminoma cells. Int J Androl. 2012 Apr 23. doi: 10.1111/j.1365‐
2605.2012.01267.x. [Epub ahead of print] IF 3.71 2