Cover Image

Spermatogenesis after Transplantation of Adipose Tissue-Derived Stem Cells in Azoospermic Guinea Pigs: A Histological and Histomorphometric Study

Mehrdokht Hajihoseini, Davood Mehrabani, Akbar Vahdati, Seyed Ebrahim Hosseini, Amin Tamadon, Mehdi Dianatpour, Farhad Rahamanifar

Background: The purpose of this research was to determine histomorphometric changes in busulfan-induced azoospermia after transplantation of AdSCs in guinea pig. AdSCs were isolated from adipose tissue around the testes of guinea pigs and characterized for mesenchymal properties.

Materials and Methods: Guinea pigs were allocated into three groups, including the control group without any intervention. To induce azoospermia, groups 2 and 3 received a dose of 40 mg/kg of busulfan with 21 days interval. Group 3 received 1×106 AdSCs in their seminiferous tubules of left testes, 35 days following last busulfan injection, while right testis in the group was considered for comparison as controls. Sixty days following transplantation of cell, histomorphometric and histopathologic changes of the experiments were assessed.

Results: After AdSCs’ transplantation, normal spermatogenesis appearance was noticed compared to busulfan-induced azoospermia and AdSCs recovered spermatogenesis, and our findings can be added to the literature in treating azoospermic infertilities.

Conclusion: The transplanted AdSCs could induce production of germinal cells using testicular seminiferous tubules and were an effective source in treating azoospermia.[GMJ.2018;7:e1000]

Adipose Tissue; Stem Cell; Azoospermia; Infertility; Busulfan; Guinea Pig

Miyamoto T, Minase G, Okabe K, Ueda H, Sengoku K. Male infertility and its genetic causes. J Obstet Gynaecol Res. 2015;41(10):1501-05. PMid:26178295

Aziz N. The importance of semen analysis in the context of azoospermia. Clinics. 2013;68:35-8.

Esteves SC, Agarwai A. The azoospermic male: current knowledge and future perspectives. Clinics. 2013;68:01-04.

Wosnitzer M, Goldstein M, Hardy MP. Review of azoospermia. Spermatogenesis. 2014;4(1):e28218.

PMid:25105055 PMCid:PMC4124057

Practice Committee of the American Society for Reproductive Medicine in collaboration with the Society for Male Reproduction and Urology. The management of infertility due to obstructive azoospermia. Fertil Steril. 2008;90(5):S121-4.


Jow WW, Steckel J, Schlegel PN, Magid MS, Goldstein M. Motile sperm in human testis biopsy specimens. J Androl. 1993;14(3):194-8.


Jarow JP, Espeland MA, Lipshultz LI. Evaluation of the azoospermic patient. J Urol. 1989;142(1):62-5.

Castilla JA, Alvarez C, Aguilar J, González-Varea C, Gonzalvo MC, Martinez L. Influence of analytical and biological variation on the clinical interpretation of seminal parameters. Hum Reprod. 2005;21(4):847-51.


Keel BA. Within-and between-subject variation in semen parameters in infertile men and normal semen donors. Fertil Steril. 2006;85(1):128-34.


Matzuk MM, Lamb DJ. Genetic dissection of mammalian fertility pathways. Nat Cell Biol. 2002;4 Suppl:s41-9.


Esteves SC. Clinical management of infertile men with nonobstructive azoospermia. Asian J Androl. 2015;17(3):459.

Venkatesan V, Madhira SL. Promise(s) of using mesenchymal stem cells in reproductive disorders. Indian J Med Res. 2014;140(Suppl 1):S98-S105.

PMid:25673552 PMCid:PMC4345762

Panahi M, Karimaghai N, Rahmanifar F, Tamadon A, Vahdati A, Mehrabani D, et al. Stereological evaluation of testes in busulfan-induced infertility of hamster. Comp Clin Pathol. 2014;24(5):1051-6.

Tamadon A, Mehrabani D, Rahmanifar F, Raayat Jahromi A, Panahi M, Zare S, et al. Induction of spermatogenesis by bone marrow-derived mesenchymal stem cells in busulfan-induced azoospermia in hamster. Int J Stem Cells. 2015;8:134-45.

PMid:26634062 PMCid:PMC4651278

Aliborzi G, Vahdati A, Mehrabani D, Ebrahim Hosseini S, Tamadon A. Isolation, characterization and growth kinetic comparison of bone marrow and adipose tissue mesenchymal stem cells of Guinea pig. Int J Stem Cells. 2015:In press.

Mehrabani D, Jaberi FM, Zakerinia M, Hadianfard MJ, Jalli R, Tanideh N, et al. The healing effect of bone marrow-derived stem cells in knee osteoarthritis: a case report. World J Plast Surg. 2016;5(2):168.

PMid:27579273 PMCid:PMC5003953

Faramarzi H, Mehrabani D, Fard M, Akhavan M, Zare S, Bakhshalizadeh S, et al. The potential of menstrual blood-derived stem cells in differentiation to epidermal lineage: A preliminary report. World J Plast Surg. 2016;5(1):26-31.

PMid:27308237 PMCid:PMC4904135

Mehrabani D, Rabiee M, Tamadon A, Zare S, Jahromi IR, Dianatpour M, et al. The growth kinetic, differentiation properties, karyotyping, and characterization of adipose tissue-derived stem cells in hamster. Comp Clin Pathol. 2016;25(5):1017-22.

Mehrabani D, Mahdiyar P, Torabi K, Robati R, Zare S, Dianatpour M, et al. Growth kinetics and characterization of human dental pulp stem cells: Comparison between third molar and first premolar teeth. J Clin Exp Dent. 2017;9(2):e172.

Mehrabani D, Rahmanifar F, Mellinejad M, Tamadon A, Dianatpour M, Zare S, et al. Isolation, culture, characterization, and adipogenic differentiation of heifer endometrial mesenchymal stem cells. Comp Clin Pathol. 2015;24(5):1159-64.

Razeghian Jahromi I, Mehrabani D, Mohammadi A, Dianatpour M, Tamadon A, Zare S, et al. The effect of fetal rat brain extract on morphology of bone marrow-derived mesenchymal stem cells. Comp Clin Pathol. 2016;25(2):343-9.

Rahmanifar F, Tamadon A, Mehrabani D, Zare S, Abasi S, Keshavarz S, et al. Histomorphometric evaluation of treatment of rat azoospermic seminiferous tubules by allotransplantation of bone marrow-derived mesenchymal stem cells. Iran J Basic Med Sci . 2016;19(6):653-61.

PMid:27482347 PMCid:PMC4951605

Mehrabani D, Hassanshahi MA, Tamadon A, Zare S, Keshavarz S, Rahmanifar F, et al. Adipose tissue-derived mesenchymal stem cells repair germinal cells of seminiferous tubules of busulfan-induced azoospermic rats. J Hum Reprod Sci. 2015;8(2):103-10.

PMid:26157302 PMCid:PMC4477447

Mehrabani D, Mehrabani G, Zare S, Manafi A. Adipose-derived stem cells (ADSC) and aesthetic surgery: a mini review. World J Plast Surg. 2013;2(2):65-70.

PMid:25489507 PMCid:PMC4238346

Hajihoseini M, Vahdati A, Ebrahim Hosseini S, Mehrabani D, Tamadon A. Induction of spermatogenesis after stem cell therapy of azoospermic guinea pigs. Vet Arh. 2017;87(3):333-50.

Noller DW, Flickinger CJ, Howards SS. Duration of the cycle of the seminiferous epithelium in the guinea pig determined by tritiated thymidine autoradiography. Biol Reprod. 1977;17(4):532-4.


Rodríguez-Casuriaga R, Geisinger A, Santi-aque FF, López-Carro B, Folle GA. High-purity flow sorting of early meiocytes based on DNA analysis of guinea pig spermatogenic cells. Cytometry A. 2011;79A(8):625-34.


Honaramooz A, Behboodi E, Hausler CL, Blash S, Ayres S, Azuma C, et al. Depletion of endogenous germ cells in male pigs and goats in preparation for germ cell transplantation. J Androl. 2005;26(6):698-705.

PMid:16291964 PMCid:PMC1352318

de Rooij DG, Vergouwen R. The estimation of damage to testicular cell lineages. Prog Clin Biol Res. 1990;372:467-80.

Berookhim BM, Schlegel PN. Azoospermia due to spermatogenic failure. Urol Clin North Am. 2014;41(1):97-113.


Cakici C, Buyrukcu B, Duruksu G, Haliloglu AH, Aksoy A, Isık A, et al. Recovery of fertility in azoospermia rats after injection of adipose-tissue-derived mesenchymal stem cells: the sperm generation. BioMed Res Int. 2013;2013:529589.

PMid:23509736 PMCid:PMC3590610

Lue Y, Erkkila K, Liu PY, Ma K, Wang C, Hikim AS, et al. Fate of bone marrow stem cells transplanted into the testis: potential implication for men with testicular failure. Am J Pathol. 2007;170(3):899-908.

PMid:17322375 PMCid:PMC1864883

Zahkook SA, Atwa A, Shahat M, Mansour AM, Bakry S. Mesenchymal stem cells restore fertility in induced azoospermic rats following chemotherapy administration. J Reprod Infertil. 2014;5(2):50-7.

Zhang D, Liu X, Peng J, He D, Lin T, Zhu J, et al. Potential spermatogenesis recovery with bone marrow mesenchymal stem cells in an azoospermic rat model. Int J Mol Sci. 2014;15(8):13151-65.

PMid:25062349 PMCid:PMC4159785

Monsefi M, Fereydouni B, Rohani L, Talaei T. Mesenchymal stem cells repair germinal cells of seminiferous tubules of sterile rats. Iran J Reprod Med. 2013;11(7):537-44.

PMid:24639788 PMCid:PMC3941348

Sabbaghi MA, Bahrami AR, Feizzade B, Kalantar SM, Matin MM, Kalantari M, et al. Trial evaluation of bone marrow derived mesenchymal stem cells (MSCs) transplantation in revival of spermatogenesis in testicular torsion. Middle East Fertil Soc J. 2012;17(4):243-49.

Drusenheimer N, Wulf G, Nolte J, Lee JH, Dev A, Dressel R, et al. Putative human male germ cells from bone marrow stem cells. Soc Reprod Fertil Suppl. 2007;63:69-76.


Nayernia K, Lee JH, Drusenheimer N, Nolte J, Wulf G, Dressel R, et al. Derivation of male germ cells from bone marrow stem cells. Lab Invest. 2006;86(7):654-63.


Chen H, Tang QL, Wu XY, Xie LC, Lin LM, Ho GY, et al. Differentiation of human umbilical cord mesenchymal stem cells into germ-like cells in mouse seminiferous tubules. Mol Med Rep. 2015;12(1):819-28.

PMid:25815600 PMCid:PMC4438948

Mital P, Kaur G, Dufour JM. Immunoprotective sertoli cells: making allogeneic and xenogeneic transplantation feasible. Reproduction. 2010;139(3):495-504.


Barry FP, Murphy JM. Mesenchymal stem cells: clinical applications and biological characterization. Int J Biochem Cell Biol. 2004;36(4):568-84.


Aghamir SMK, Salavati A, Yousefie R, Tootian Z, Ghazaleh N, Jamali M, et al. Does bone marrow-derived mesenchymal stem cell transfusion prevent antisperm antibody production after traumatic testis rupture? Urology. 2014;84(1):82-6.


Meinhardt A, Hedger MP. Immunological, paracrine and endocrine aspects of testicular immune privilege. Mol Cell Endocrinol. 2011;335(1):60-8.


Suttorp M, Millot F. Treatment of pediatric chronic myeloid leukemia in the year 2010: use of tyrosine kinase inhibitors and stem-cell transplantation. Hematology Am Soc Hematol Educ Program. 2010;2010(1):368-76.

Bartelink I, van Reij E, Gerhardt C, van Maarseveen E, de Wildt A, Versluys B, et al. Fludarabine and exposure-targeted busulfan compares favorably with busulfan/cyclophosphamide-based regimens in pediatric hematopoietic cell transplantation: maintaining efficacy with less toxicity. Biol Blood Marrow Transplant. 2014;20(3):345-53.



  • There are currently no refbacks.