{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T09:50:16Z","timestamp":1790502616138,"version":"4.1.0"},"reference-count":35,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2021,2,26]],"date-time":"2021-02-26T00:00:00Z","timestamp":1614297600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003977","name":"Israel Science Foundation","doi-asserted-by":"publisher","award":["295\/14"],"award-info":[{"award-number":["295\/14"]}],"id":[{"id":"10.13039\/501100003977","id-type":"DOI","asserted-by":"publisher"}]},{"name":"The REPRODUCTION HUB, Faculty of Health Sciences, Ben-Gurion University of the Negev","award":["111"],"award-info":[{"award-number":["111"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Spermatogenesis is a complex process, in which spermatogonial cells proliferate and differentiate in the seminiferous tubules of the testis to generate sperm. This process is under the regulation of endocrine and testicular paracrine\/autocrine factors. In the present study, we demonstrated that colony stimulating factor-1 (CSF-1) is produced by mouse testicular macrophages, Leydig, Sertoli, peritubular cells and spermatogonial cells (such as CDH1-positively stained cells; a marker of spermatogonial cells). In addition, we demonstrated the presence of CSF-1 and its receptor (CSF-1R) in testicular macrophages, Leydig, Sertoli, peritubular cells and spermatogonial cells of human testis. We also show that the protein levels of CSF-1 were the highest in testis of 1-week-old mice and significantly decreased with age (2\u201312-week-old). However, the transcriptome levels of CSF-1 significantly increased in 2\u20133-week-old compared to 1-week-old, and thereafter significantly decreased with age. On the other hand, the transcriptome levels of CSF-1R was significantly higher in mouse testicular tissue of all examined ages (2\u201312-week-old) compared to 1-week-old. Our results demonstrate the involvement of CSF-1 in the induction the proliferation and differentiation of spermatogonial cells to meiotic and postmeiotic stages (BOULE- and ACROSIN-positive cells) under in vitro culture conditions, using methylcellulose culture system (MCS). Thus, it is possible to suggest that CSF-1 system, as a testicular paracrine\/autocrine system, is involved in the development of different stages of spermatogenesis and may be used in the development of future therapeutic strategies for treatment of male infertility.<\/jats:p>","DOI":"10.3390\/ijms22052325","type":"journal-article","created":{"date-parts":[[2021,2,26]],"date-time":"2021-02-26T04:36:24Z","timestamp":1614314184000},"page":"2325","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["The Presence of Colony-Stimulating Factor-1 and Its Receptor in Different Cells of the Testis; It Involved in the Development of Spermatogenesis In Vitro"],"prefix":"10.3390","volume":"22","author":[{"given":"Alaa","family":"Sawaied","sequence":"first","affiliation":[{"name":"The Shraga Segal Department of Microbiology, Immunology, and Genetics, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva 8410501, Israel"},{"name":"The Center of Advanced Research and Education in Reproduction (CARER), Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva 8410501, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eden","family":"Arazi","sequence":"additional","affiliation":[{"name":"The Shraga Segal Department of Microbiology, Immunology, and Genetics, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva 8410501, Israel"},{"name":"The Center of Advanced Research and Education in Reproduction (CARER), Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva 8410501, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahmad","family":"AbuElhija","sequence":"additional","affiliation":[{"name":"The Shraga Segal Department of Microbiology, Immunology, and Genetics, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva 8410501, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eitan","family":"Lunenfeld","sequence":"additional","affiliation":[{"name":"The Center of Advanced Research and Education in Reproduction (CARER), Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva 8410501, Israel"},{"name":"Department of OB\/GYN, Soroka Medical Center, Beer Sheva 8410501, Israel"},{"name":"Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva 8410501, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4246-9299","authenticated-orcid":false,"given":"Mahmoud","family":"Huleihel","sequence":"additional","affiliation":[{"name":"The Shraga Segal Department of Microbiology, Immunology, and Genetics, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva 8410501, Israel"},{"name":"The Center of Advanced Research and Education in Reproduction (CARER), Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva 8410501, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,2,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"628","DOI":"10.1016\/j.tcb.2004.09.016","article-title":"CSF-1 regulation of the wandering macrophage: Complexity in action","volume":"14","author":"Pixley","year":"2004","journal-title":"Trends Cell Biol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1107","DOI":"10.1016\/j.celrep.2015.07.015","article-title":"Macrophages Contribute to the Spermatogonial Niche in the Adult Testis","volume":"12","author":"DeFalco","year":"2015","journal-title":"Cell Rep."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1191","DOI":"10.1242\/dev.032243","article-title":"Colony stimulating factor 1 is an extrinsic stimulator of mouse spermatogonial stem cell self-renewal","volume":"136","author":"Oatley","year":"2009","journal-title":"Development"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"5128","DOI":"10.4049\/jimmunol.154.10.5128","article-title":"Colony-stimulating factor-1 secreted by fibroblasts promotes the growth of dendritic cell lines (XS series) derived from murine epidermis","volume":"154","author":"Takashima","year":"1995","journal-title":"J. Immunol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"616","DOI":"10.1002\/(SICI)1097-4547(19990901)57:5<616::AID-JNR4>3.0.CO;2-E","article-title":"Expression of colony stimulating factor-1 receptor (CSF-1R) by CNS neurons in mice","volume":"57","author":"Wang","year":"1999","journal-title":"J. Neurosci. Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"848","DOI":"10.1083\/jcb.91.3.848","article-title":"Distribution of cells bearing receptors for a colony-stimulating factor (csf-1) in murine tissues","volume":"91","author":"Byrne","year":"1981","journal-title":"J. Cell Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1083\/jcb.85.1.153","article-title":"Specific interaction of murine colony-stimulating factor with mononuclear phagocytic cells","volume":"85","author":"Guilbert","year":"1980","journal-title":"J. Cell Biol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1016\/S8756-3282(95)00163-8","article-title":"Detection of transcripts and binding sites for colony-stimulating factor-1 during bone development","volume":"17","author":"Hofstetter","year":"1995","journal-title":"Bone"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"8818","DOI":"10.1073\/pnas.86.22.8818","article-title":"Temporal expression and location of colony-stimulating factor 1 (CSF-I) and its receptor in the female reproductive tract are consistent with CSF-1-regulated placental development","volume":"86","author":"Arceci","year":"1989","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1016\/0012-1606(89)90319-9","article-title":"Expression of the c-fms proto-oncogene and of the cytokine, CSF-1, during mouse embryogenesis","volume":"133","author":"Regenstreif","year":"1989","journal-title":"Dev. Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1196\/annals.1411.014","article-title":"Identification of testis-specific male contraceptive targets: Insights from transcriptional profiling of the cycle of the rat seminiferous epithelium and purified testicular cells","volume":"1120","author":"Johnston","year":"2007","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1095\/biolreprod.108.073809","article-title":"The molecular signature of spermatogonial stem\/progenitor cells in the 6-day-old mouse testis","volume":"80","author":"Kokkinaki","year":"2009","journal-title":"Biol. Reprod."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Sawaied, A., Lunenfeld, E., and Huleihel, M. (2020). Interleukin-34, a Novel Paracrine\/Autocrine Factor in Mouse Testis, and Its Possible Role in the Development of Spermatogonial Cells In Vitro. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21218143"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1636","DOI":"10.1210\/mend.11.11.0009","article-title":"Colony-Stimulating Factor-1 Plays a Major Role in the Development of Reproductive Function in Male Mice","volume":"11","author":"Cohen","year":"1997","journal-title":"Mol. Endocrin."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1095\/biolreprod55.2.310","article-title":"Absence of Colony-Stimulating Factor-1 in Osteopetrotic (csfmoP\/csfmOP) Mice Results in Male Fertility Defects","volume":"55","author":"Cohen","year":"1996","journal-title":"Biol. Reprod."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1290","DOI":"10.1095\/biolreprod56.5.1290","article-title":"Effect of colony stimulating factor-1 (CSF-1) null mutation, osteopetrotic (csfmop), on the distribution of macrophages in the male reproductive tract","volume":"56","author":"Pollard","year":"1997","journal-title":"Biol. Reprod."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2355","DOI":"10.1002\/stem.1238","article-title":"Concise review: Spermatogenesis in an artificial three-dimensional system","volume":"30","author":"Mahmoud","year":"2012","journal-title":"Stem Cells"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"972","DOI":"10.4103\/1008-682X.154994","article-title":"Application of three-dimensional culture systems to study mammalian spermatogenesis, with an emphasis on the rhesus monkey (Macaca mulatta)","volume":"17","author":"Huleihel","year":"2015","journal-title":"Asian J. Aandrol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.scr.2018.04.009","article-title":"Spermatogonial stem cells and spermatogenesis in mice, monkeys and men","volume":"29","author":"Fayomi","year":"2018","journal-title":"Stem Cell Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1038\/nrn812","article-title":"The GDNF family: Signalling, biological functions and therapeutic value","volume":"3","author":"Airaksinen","year":"2002","journal-title":"Nat. Rev. Neuro."},{"key":"ref_21","first-page":"259","article-title":"Regulation of spermatogenesis by paracrine\/autocrine testicular factors","volume":"6","author":"Huleihel","year":"2004","journal-title":"Asian J. Androl."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1111\/andr.12165","article-title":"The Sertoli cell: One hundred fifty years of beauty and plasticity","volume":"4","author":"Hess","year":"2016","journal-title":"Andrology"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1038\/aja.2011.112","article-title":"Differentiation of murine male germ cells to spermatozoa in a soft agar culture system","volume":"14","author":"Elhija","year":"2012","journal-title":"Asian J. Androl."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1093\/molehr\/gap052","article-title":"New horizons for in vitro spermatogenesis? An update on novel three-dimensional culture systems as tools for meiotic and post-meiotic differentiation of testicular germ cells","volume":"15","author":"Stukenborg","year":"2009","journal-title":"Mol. Hum. Reprod."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1007","DOI":"10.1089\/scd.2017.0301","article-title":"Development of postmeiotic cells in vitro from spermatogonial cells of prepubertal cancer patients","volume":"27","author":"AbuMadighem","year":"2018","journal-title":"Stem Cells Dev."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"AbuMadighem, A., Solomon, R., Stepanovsky, A., Kapelushnik, J., Shi, Q., Meese, E., Lunenfeld, E., and Huleihel, M. (2018). Development of spermatogenesis in vitro in three-dimensional culture from spermatogonial cells of busulfan-treated immature mice. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19123804"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Abofoul-Azab, M., Lunenfeld, E., Levitas, E., Zeadna, A., Younis, J.S., Bar-Ami, S., and Huleihel, M. (2019). Identification of Premeiotic, Meiotic, and Postmeiotic Cells in Testicular Biopsies without Sperm from Sertoli Cell-Only Syndrome Patients. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20030470"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1182\/blood.V99.1.111","article-title":"Targeted disruption of the mouse colony-stimulating factor 1 receptor gene results in osteopetrosis, mononuclear phagocyte deficiency, increased primitive progenitor cell frequencies, and reproductive defects","volume":"99","author":"Dai","year":"2002","journal-title":"Blood"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1189\/jlb.2A0114-006R","article-title":"Pleiotropic effects of extended blockade of CSF1R signaling in adult mice","volume":"96","author":"Sauter","year":"2014","journal-title":"J. Leukoc. Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"765","DOI":"10.1002\/jlb.66.5.765","article-title":"Macrophages: Important accessory cells for reproductive function","volume":"66","author":"Cohen","year":"1999","journal-title":"J. Leukoc. Biol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1177\/153537020623100101","article-title":"Physiologic interactions between macrophages and Leydig cells","volume":"231","author":"Hutson","year":"2006","journal-title":"Exp. Biol. Med."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"4828","DOI":"10.1073\/pnas.87.12.4828","article-title":"Total absence of colony-stimulating factor 1 in the macrophage-deficient osteopetrotic (op\/op) mouse","volume":"87","author":"Bartocci","year":"1990","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1016\/j.diff.2013.06.005","article-title":"Glial cell line-derived neurotrophic factor (GDNF) induced migration of spermatogonial cells in vitro via MEK and NF-kB pathways","volume":"86","author":"Huleihel","year":"2013","journal-title":"Differentiation"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2\u2212\u2206\u2206Ct Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1136\/jmedgenet-2017-104514","article-title":"Mutation in TDRD9 causes non-obstructive azoospermia in infertile men","volume":"54","author":"Arafat","year":"2017","journal-title":"J. Med. Genet."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/5\/2325\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:28:59Z","timestamp":1760160539000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/5\/2325"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,2,26]]},"references-count":35,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2021,3]]}},"alternative-id":["ijms22052325"],"URL":"https:\/\/doi.org\/10.3390\/ijms22052325","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,2,26]]}}}