BACKGROUND: Inflammation-induced testicular damage is a significant contributing factor to the increasing incidence of infertility. Traditional treatments during the inflammatory phase often fail to achieve the desired fertility outcomes, necessitating innovative interventions such as cell therapy. METHODS: We explored the in vivo properties of intravenously administered Sertoli cells (SCs) in an acute lipopolysaccharide (LPS)-induced inflammatory mouse model. Infiltrating and resident myeloid cell phenotypes were assessed using flow cytometry. The impact of SC administration on testis morphology and germ cell quality was evaluated using computer-assisted sperm analysis (CASA) and immunohistochemistry. RESULTS: SCs demonstrated a distinctive migration pattern, importantly they preferentially concentrated in the testes and liver. SC application significantly reduced neutrophil infiltration as well as preserved the resident macrophage subpopulations. SCs upregulated MerTK expression in both interstitial and peritubular macrophages. Applied SC treatment exhibited protective effects on sperm including their motility and kinematic parameters, and maintained the physiological testicular morphology. CONCLUSION: Our study provides compelling evidence of the therapeutic efficacy of SC transplantation in alleviating acute inflammation-induced testicular damage. These findings contribute to the expanding knowledge on the potential applications of cell-based therapies for addressing reproductive health challenges and offer a promising approach for targeted interventions in male infertility.

Department of Cell Biology Faculty of Science Charles University Vinicna 7 Prague 2 128 00 Czech Republic

Department of Physiology Faculty of Science Charles University Vinicna 7 Prague 2 128 00 Czech Republic

Department of Zoology Faculty of Science Charles University Vinicna 7 Prague 2 128 00 Czech Republic

Laboratory of Reproductive Biology Institute of Biotechnology of the Czech Academy of Sciences BIOCEV Vestec Prumyslova 595 Prague 252 50 Czech Republic

Zobrazit více v PubMed

Sharlip ID, Jarow JP, Belker AM, Lipshultz LI, Sigman M, Thomas AJ, et al. Best practice policies for male infertility. Fertil Steril. 2002;77:873–82. PubMed

Fijak M, Pilatz A, Hedger MP, Nicolas N, Bhushan S, Michel V, et al. Infectious, inflammatory and ‘autoimmune’ male factor infertility: how do rodent models inform clinical practice? Hum Reprod Update. 2018;24:416–41. PubMed PMC

Gallegos-Avila G, Ortega-Martínez M, Ramos-González B, Tijerina-Menchaca R, Ancer-Rodríguez J, Jaramillo-Rangel G. Ultrastructural findings in semen samples of infertile men infected with Chlamydia trachomatis and mycoplasmas. Fertil Steril. 2009;91:915–9. PubMed

Sobinoff AP, Dando SJ, Redgrove KA, Sutherland JM, Stanger SJ, Armitage CW, et al. Chlamydia muridarum infection-induced destruction of male germ cells and sertoli cells is partially prevented by Chlamydia major outer membrane protein-specific immune CD4 cells. Biol Reprod. 2015;92:27. PubMed

Bryan ER, Kim J, Beagley KW, Carey AJ. Testicular inflammation and infertility: could chlamydial infections be contributing? Am J Reprod Immunol. 2020;84:e13286. PubMed

Henkel R, Offor U, Fisher D. The role of infections and leukocytes in male infertility. Andrologia. 2021;53:e13743. PubMed

Liakath Ali F, Park HS, Beckman A, Eddy AC, Alkhrait S, Ghasroldasht MM, et al. Fertility Protection, a Novel Concept: umbilical cord mesenchymal stem cell-derived exosomes protect against Chemotherapy-Induced Testicular cytotoxicity. Int J Mol Sci. 2024;25:60. PubMed PMC

Calogero AE, Condorelli RA, Russo GI, Vignera S la. Conservative nonhormonal options for the treatment of male infertility: Antibiotics, anti-inflammatory drugs, and antioxidants. Biomed Res Int. 2017;2017:4650182. PubMed PMC

Behnke J, Kremer S, Shahzad T, Chao CM, Böttcher-Friebertshäuser E, Morty RE, et al. MSC Based therapies—New perspectives for the injured lung. J Clin Med. 2020;9:682. PubMed PMC

Jasim SA, Yumashev AV, Abdelbasset WK, Margiana R, Markov A, Suksatan W, et al. Shining the light on clinical application of mesenchymal stem cell therapy in autoimmune diseases. Stem Cell Res Ther. 2022;13:1–15. PubMed PMC

Kadam P, Ntemou E, Baert Y, Van Laere S, Van Saen D, Goossens E. Co-transplantation of mesenchymal stem cells improves spermatogonial stem cell transplantation efficiency in mice. Stem Cell Res Ther. 2018;9:317. PubMed PMC

Hsiao CH, Ji ATQ, Chang CC, Cheng CJ, Lee LM, Ho JHC. Local injection of mesenchymal stem cells protects testicular torsion-induced germ cell injury Rocky Tuan; Timothy O’Brien. Stem Cell Res Ther. 2015;6:113. PubMed PMC

Griswold MD. 50 years of spermatogenesis: sertoli cells and their interactions with germ cells. Biol Reprod. 2018;99:87. PubMed PMC

Kaur G, Thompson LA, Dufour JM. Sertoli cells–immunological sentinels of spermatogenesis. Semin Cell Dev Biol. 2014;30:36–44. PubMed PMC

Kamińska A, Pardyak L, Lustofin S, Gielata K, Arent Z, Pietsch-Fulbiszewska A, et al. 9-cis-retinoic acid signaling in sertoli cells regulates their immunomodulatory function to control lymphocyte physiology and Treg differentiation. Reprod Biol Endocrinol. 2024;22:75. PubMed PMC

Washburn RL, Hibler T, Kaur G, Dufour JM. Sertoli cell Immune Regulation: a double-edged Sword. Front Immunol. 2022;13:913502. PubMed PMC

Chiappalupi S, Salvadori L, Borghi M, Mancuso F, Pariano M, Riuzzi F, et al. Grafted sertoli cells exert Immunomodulatory Non-immunosuppressive effects in Preclinical models of infection and Cancer. Cells. 2024;13:544. PubMed PMC

Shinohara T, Orwig KE, Avarbock MR, Brinster RL. Restoration of Spermatogenesis in Infertile mice by sertoli cell transplantation. Biol Reprod. 2003;68:1064–71. PubMed

Zhang Z, Shao S, Shetty G, Meistrich ML. Donor sertoli cells transplanted into irradiated rat testes stimulate partial recovery of endogenous spermatogenesis. Reproduction. 2009;137:497–508. PubMed

Washburn RL, Hibler T, Thompson LA, Kaur G, Dufour JM. Therapeutic application of sertoli cells for treatment of various diseases. Semin Cell Dev Biol. 2022;121:10–23. PubMed

Campese AF, Grazioli P, de Cesaris P, Riccioli A, Bellavia D, Pelullo M, et al. Mouse sertoli cells sustain de novo generation of regulatory T cells by triggering the notch pathway through soluble JAGGED1. Biol Reprod. 2014;90:53. PubMed

Gao J, Wang X, Wang Y, Han F, Cai W, Zhao B, et al. Murine sertoli cells promote the development of tolerogenic dendritic cells: a pivotal role of galectin-1. Immunology. 2016;148:253–65. PubMed PMC

Chen R, Wang F, Chen Y, Han D. Immune homeostasis and disorder in the testis – roles of sertoli cells. J Reprod Immunol. 2022;152:103625. PubMed

Porubska B, Vasek D, Somova V, Hajkova M, Hlaviznova M, Tlapakova T, et al. Sertoli Cells Possess Immunomodulatory Properties and the ability of mitochondrial transfer similar to mesenchymal stromal cells. Stem Cell Rev Rep. 2021;17:1905–16. PubMed

Bhushan S, Theas MS, Guazzone VA, Jacobo P, Wang M, Fijak M, et al. Immune Cell subtypes and their function in the Testis. Front Immunol. 2020;11:583304. PubMed PMC

Lang V, Ferencik S, Ananthasubramaniam B, Kramer A, Maier B. Susceptibility rhythm to bacterial endotoxin in myeloid clock-knockout mice. Elife. 2021;10:e62469. PubMed PMC

Bilinska B, Hejmej A, Kotula-Balak M. Preparation of testicular samples for histology and immunohistochemistry. Methods Mol Biol. 2018;1748:17–36. PubMed

Nguyen TMX, Vegrichtova M, Tlapakova T, Krulova M, Krylov V. The interconnection between cytokeratin and cell membrane-bound β-catenin in sertoli cells derived from juvenile Xenopus tropicalis testes. Biol Open. 2019;8:bio043950. PubMed PMC

Hedger MP. Immunophysiology and Pathology of inflammation in the Testis and Epididymis. J Androl. 2011;32:625. PubMed PMC

Hajkova M, Javorkova E, Zajicova A, Trosan P, Holan V, Krulova M. A local application of mesenchymal stem cells and cyclosporine a attenuates immune response by a switch in macrophage phenotype. J Tissue Eng Regen Med. 2017;11:1456–65. PubMed

Bhushan S, Meinhardt A. The macrophages in testis function. J Reprod Immunol. 2017;119:107–12. PubMed

Indumathy S, Pueschl D, Klein B, Fietz D, Bergmann M, Schuppe HC, et al. Testicular immune cell populations and macrophage polarisation in adult male mice and the influence of altered activin A levels. J Reprod Immunol. 2020;142:103204. PubMed

Mossadegh-Keller N, Gentek R, Gimenez G, Bigot S, Mailfert S, Sieweke MH. Developmental origin and maintenance of distinct testicular macrophage populations. J Exp Med. 2017;214:2829. PubMed PMC

Lindsay RS, Whitesell JC, Dew KE, Rodriguez E, Sandor AM, Tracy D, et al. MERTK on mononuclear phagocytes regulates T cell antigen recognition at autoimmune and tumor sites. J Exp Med. 2021;218:e20200464. PubMed PMC

Shi J, Gao S, Chen Z, Chen Z, Yun D, Wu X, et al. Absence of MerTK disrupts spermatogenesis in an age-dependent manner. Mol Cell Endocrinol. 2023;560:111815. PubMed

Gu X, Li SY, DeFalco T. Immune and vascular contributions to organogenesis of the testis and ovary. FEBS J. 2022;289:2386. PubMed PMC

Azenabor A, Ekun AO, Akinloye O. Impact of inflammation on male Reproductive Tract. J Reprod Infertil. 2015;16:123–9. PubMed PMC

Mastrolia I, Foppiani EM, Murgia A, Candini O, Samarelli AV, Grisendi G, et al. Challenges in Clinical Development of Mesenchymal Stromal/Stem cells: Concise Review. Stem Cells Transl Med. 2019;8:1135–48. PubMed PMC

Li J, Wu Z, Zhao L, Liu Y, Su Y, Gong X et al. The heterogeneity of mesenchymal stem cells: an important issue to be addressed in cell therapy. Stem Cell Res Ther. 2023;14. PubMed PMC

Gong D, Zhang C, Li T, Zhang J, Zhang N, Tao Z, et al. Are sertoli cells a kind of mesenchymal stem cells? Am J Transl Res. 2017;9:1067–74. PubMed PMC

Kurtz A. Mesenchymal stem cell delivery routes and fate. Int J Stem Cells. 2008;1:1–7. PubMed PMC

Zhong L, Yang M, Zou X, Du T, Xu H, Sun J. Human umbilical cord multipotent mesenchymal stromal cells alleviate acute ischemia-reperfusion injury of spermatogenic cells via reducing inflammatory response and oxidative stress. Stem Cell Res Ther. 2020;11:294. PubMed PMC

Moritoki Y, Kojima Y, Mizuno K, Kamisawa H, Kohri K, Hayashi Y. Intratesticular pressure after testicular torsion as a predictor of subsequent spermatogenesis: a rat model. BJU Int. 2012;109:466–70. PubMed

Winnall WR, Muir JA, Hedger MP. Rat resident testicular macrophages have an alternatively activated phenotype and constitutively produce interleukin-10 in vitro. J Leukoc Biol. 2011;90:133–43. PubMed

Celebi M, Paul AGA. Blocking E-selectin inhibits ischaemia-reperfusion-induced neutrophil recruitment to the murine testis. Andrologia. 2008;40:235–9. PubMed

Bouchery T, Harris N. Neutrophil–macrophage cooperation and its impact on tissue repair. Immunol Cell Biol. 2019;97:289–98. PubMed

Barman PK, Shin JE, Lewis SA, Kang S, Wu D, Wang Y, et al. Production of MHCII-expressing classical monocytes increases during aging in mice and humans. Aging Cell. 2022;21:e13701. PubMed PMC

Guilliams M, Mildner A, Yona S. Developmental and Functional Heterogeneity of monocytes. Immunity. 2018;49:595–613. PubMed

Bain CC, Bravo-Blas A, Scott CL, Gomez Perdiguero E, Geissmann F, Henri S, et al. Constant replenishment from circulating monocytes maintains the macrophage pool in adult intestine. Nat Immunol. 2014;15:929. PubMed PMC

Wright PB, McDonald E, Bravo-Blas A, Baer HM, Heawood A, Bain CC, et al. The mannose receptor (CD206) identifies a population of colonic macrophages in health and inflammatory bowel disease. Sci Rep. 2021;11:19616. PubMed PMC

Gu X, Li SY, DeFalco T. Immune and vascular contributions to organogenesis of the testis and ovary. FEBS J. 2022;289:2386–408. PubMed PMC

Wanjari UR, Gopalakrishnan AV. A review on immunological aspects in male reproduction: an immune cells and cytokines. J Reprod Immunol. 2023;158:103984. PubMed

Hayrabedyan S, Todorova K, Jabeen A, Metodieva G, Toshkov S, Metodiev MV, et al. Sertoli cells have a functional NALP3 inflammasome that can modulate autophagy and cytokine production. Sci Rep. 2016;6:18896. PubMed PMC

Martínez-Pastor F. What is the importance of sperm subpopulations? Anim Reprod Sci. 2022;246:106844. PubMed

Ferraz MAMM, Morató R, Yeste M, Arcarons N, Pena AI, Tamargo C, et al. Evaluation of sperm subpopulation structure in relation to in vitro sperm–oocyte interaction of frozen-thawed semen from Holstein bulls. Theriogenology. 2014;81:1067–72. PubMed

Sheng ZY, Gao N, Wang ZY, Cui XY, Zhou DS, Fan DY, et al. Sertoli cells are susceptible to ZIKV infection in mouse testis. Front Cell Infect Microbiol. 2017;7:274103. PubMed PMC

Galderisi U, Peluso G, Di Bernardo G. Clinical trials based on mesenchymal stromal cells are exponentially increasing: where are we in recent years? Stem Cell Reviews Rep. 2021;1:1–14. PubMed PMC

Nasir Uddin MM, Ahmed S, Kabir MSH, Rahman MS, Sultan RA, Emran T, Bin. In vivo analgesic, anti-inflammatory potential in Swiss albino mice and in vitro thrombolytic activity of hydroalcoholic fruits extract from Daemonorops Robusta Warb. J Appl Pharm Sci. 2017;7:104–13.

Mitra S, Islam F, Das R, Urmee H, Akter A, Idris AM, et al. Pharmacological potential of Avicennia Alba Leaf Extract: an experimental analysis focusing on Antidiabetic, anti-inflammatory, analgesic, and Antidiarrheal Activity. Biomed Res Int. 2022;2022:7624189. PubMed PMC

Sakib SA, Tareq AM, Islam A, Rakib A, Islam MN, Uddin MA, et al. Anti-inflammatory, thrombolytic and hair-growth promoting activity of the n-Hexane fraction of the methanol extract of Leea indica leaves. Plants. 2021;10:1081. PubMed PMC

Luca G, Baroni T, Arato I, Hansen BC, Cameron DF, Calafiore R. Role of sertoli cell proteins in Immunomodulation. Protein Pept Lett. 2018;25:440–5. PubMed

Vegrichtova M, Hajkova M, Porubska B, Vasek D, Krylov V, Tlapakova T et al. Xenogeneic sertoli cells modulate immune response in an evolutionary distant mouse model through the production of interleukin-10 and PD-1 ligands expression. Xenotransplantation. 2022;e12742. PubMed

Matsumoto S, Matsumoto K. Clinical islet xenotransplantation: development of isolation Protocol, anti-rejection strategies, and clinical outcomes. Cells. 2024;13:828. PubMed PMC

Yin Z, Chen D, Hu F, Ruan Y, Li J, Wang L, et al. Cotransplantation with xenogenetic neonatal porcine sertoli cells significantly prolongs islet allograft survival in nonimmunosuppressive rats. Transplantation. 2009;88:339–45. PubMed

Lakpour MR, Aghajanpour S, Koruji M, Shahverdi A, Sadighi-Gilani MA, Sabbaghian M, et al. Isolation, culture and characterization of human sertoli cells by Flow Cytometry: Development of Procedure. J Reprod Infertil. 2017;18:213–7. PubMed PMC

Chui K, Trivedi A, Cheng CY, Cherbavaz DB, Dazin PF, Huynh ALT, et al. Characterization and functionality of proliferative human sertoli cells. Cell Transpl. 2011;20:619. PubMed PMC

Bistoni G, Calvitti M, Mancuso F, Arato I, Falabella G, Cucchia R, et al. Prolongation of skin allograft survival in rats by the transplantation of microencapsulated xenogeneic neonatal porcine sertoli cells. Biomaterials. 2012;33:5333–40. PubMed