Histomorphometric Evaluation of Allogeneic Transplantation of Bone Marrow Mesenchymal Stem Cells in Azoospermic Mice Model

Ahmad Mozafar, Davood Mehrabani, Akbar Vahdati, Ebrahim Hosseini, Mohsen Forouzanfar

Abstract


BACKGROUND: Stem cell-based therapy is one of the newest and evolving techniques in reproductive medicine. The aim of this study was to investigate the effect of allogeneic bone marrow mesenchymal stem cells (BM-MSCs) transplantation on the testis of busulfan induced azoospermia in Balb/C mice.

METHODS: Eighteen adult Balb/C mice were divided into three equal groups including control, busulfan and busulfan+cell therapy (busul+CT). For induction of azoospermia, busulfan and busul+CT groups received two injections of 10 mg/Kg of busulfan intraperitoneally with 21 days interval. In the cell therapy group 35 days after the last injection of busulfan, cluster of differentiation (CD)90+/CD34-/CD45- BM-MSCs were injected into the efferent duct of testis. Eight weeks after the BM-MSCs therapy, mice were sacrificed and tissues were taken for histological and histomorphometric evaluations.

RESULTS: In busul+CT group, cellular and total diameters and cellular and cross-sectional areas significantly increased in comparison to busulfan group (p˂0.001), but there were no significant differences between busul+CT and control group (p˃0.05). Numerical density and tubular count per area unit in busul+CT and control groups were significantly less than busulfan group (p˂0.001), but there were no significant difference between busul+CT and control group (p˃0.05). The luminal diameter and area showed no significant change in all groups (p˃0.05). In busul+CT group, spermatogenesis index significantly increased when compared to busulfan and control groups (p˂0.001 and p˂0.05, respectively).

CONCLOSION: Histomorphometric findings showed CD90+/CD34-/CD45- BM-MSCs transplantation on the testis of busulfan-induced azoospermic in Balb/C mice recovered spermatogenesis.

KEYWORDS: mesenchymal stem cell, cell therapy, azoospermia, busulfan, mouse


Full Text:

PDF

References


Gudeloglu A, Parekattil SJ. Update in the evaluation of the azoospermic male. Clinics (Sao Paulo). 2013; 68: 27-34, CrossRef.

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: 13151-65, CrossRef.

Vassilakopoulou M, Boostandoost E, Papaxoinis G, de La Motte Rouge T, Khayat D, Psyrri A. Anticancer treatment and fertility: Effect of therapeutic modalities on reproductive system and functions. Crit Rev Oncol Hematol. 2016; 97: 328-34, CrossRef.

Meistrich ML. Effects of chemotherapy and radiotherapy on spermatogenesis in humans. Fertil Steril. 2013; 100: 1180-6, CrossRef.

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

Cakici C, Buyrukcu B, Duruksu G, Haliloglu AH, AksoyA, Isık A, et al. Recovery of fertility in azoospermia rats after injection of adiposetissue derived mesenchymal stem cells: The sperm generation. BioMed Res Int. 2013; 2013: 1-18, CrossRef.

Asadi-Yousefabad S-L, Khodakaram-Tafti A, Dianatpour M, Mehrabani D, Zare S, Tamadon A, et al. Genetic evaluation of bone marrowderived mesenchymal stem cells by a modified karyotyping method. Comp Clin Pathol. 2015; 24: 1361-6, CrossRef.

Mahdiyar P, Zare S, Robati R, Dianatpour M, Torabi K, Tamadon A, et al. Isolation, culture, and characterization of human dental pulp mesenchymal stem cells. Int J Pediatr. 2014; 2: 44, CrossRef.

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: 103-10, CrossRef.

Ehninger A, Trumpp A. The bone marrow stem cell niche grows up: mesenchymal stem cells and macrophages move in. J Exp Med. 2011; 208: 421-28, CrossRef.

Payehdar A, Hosseini E, Mehrabani D, Forouzanfar M. Busulfan treatment effects on testicular tissue and serum levels of antimullerian hormone and testosterone in adult mice. Indones Biomed J. 2017; 9: 106-12, CrossRef.

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

Rahmanifar F, Tamadon A, Mehrabani D, Zare Sh, 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: 653-61, PMID.

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: 368-76, CrossRef.

Le Bourgeois A, Lestang E, Guillaume T, Delaunay J, Ayari S, Blin N, et al. Prognostic impact of immune status and hematopoietic recovery before and after fludarabine, IV busulfan, and antithymocyte globulins (FB2 regimen) reduced-intensity conditioning regimen (RIC) allogeneic stem cell transplantation (allo-SCT). Eur J Haematol. 2013; 90: 177-86, CrossRef.

Nieto Y, Thall P, Valdez B, Andersson B, Popat U, Anderlini P, et al. High-dose infusional gemcitabine combined with busulfan and melphalan with autologous stem-cell transplantation in patients with refractory lymphoid malignancies. Biol Blood Marrow Transplant. 2012; 18: 1677-86, CrossRef.

Iwamoto T, Hiraku Y, Oikawa S, Mizutani H, Kojima M, Kawanishi S. DNA intrastrand cross-link at the 5′-GA-3′ sequence formed by busulfan and its role in the cytotoxic effect. Cancer Sci. 2004; 95: 454-58, CrossRef.

Gutierrez K, Glanzner WG, Chemeris RO, Rigo ML, Comim FV, Bordignon V, et al. Gonadotoxic effects of busulfan in two strains of mice. Reprod Toxicol. 2016; 59: 31-9, CrossRef.

Berookhim B.M., Schlegel. Azoospermia due to spermatogenic failure. Urol Clin. North Am. 2014; 41: 97-113, CrossRef.

Caplan AI. Adult mesenchymal stem cells for tissue engineering versus regenerative medicine. J Cell Physiol. 2007; 213: 341-7, CrossRef.

Caplan AI, Dennis JE. Mesenchymal stem cells as trophic mediators. J Cell Biochem. 2006; 98: 1076-84, CrossRef.

Kuo TK, Hung SP, Chuang CH, Chen CT, Shih YR, Fang SC, et al. Stem cell therapy for liver disease: parameters governing the success of using bone marrow mesenchymal stem cells. Gastroenterology. 2008; 134: 2111-21, CrossRef.

Amado LC, Saliaris AP, Schuleri KH, St John M, Xie JS, Cattaneo S, et al. Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction. Proc Natl Acad Sci USA. 2005; 102: 11474-9, CrossRef.

Sadraie MR, Mehrabani D, Vahdari A. Comparison of therapeutic effects of bone marrow mesenchymal stem cells and liquid culture environment (secreta) in the treatment of induced knee abrasion created in Guinea Pigs. Armaghane danesh. 2016; 20: 651-65.

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

Hajihoseini H, Vahdati A, Hosseini E, Mehrabani D, Tamadon A. Induction of spermatogenesis after stem cell therapy of azoospermic guinea pigs. Vet Arhiv. 2017; 87: 333-50, CrossRef.

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: 899-908, CrossRef.

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: 537-44, PMID.

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 spermatogenesisin testicular torsion. Middle East Fertil Soc J. 2012; 17: 243-9, CrossRef.

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: 654-63, CrossRef.

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: 819-28, CrossRef.

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

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

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

Ai J, Ebrahimi S, Khoshzaban A, Jafarzadeh Kashi TS, Mehrabani D. Tissue engineering using human mineralized bone xenograft and bone marrow mesenchymal stem cells allograft in healing of tibial fracture of experimental rabbit model. Iran Red Crescent Med J. 2012; 14: 96-103, PMID.




DOI: https://doi.org/10.18585/inabj.v10i2.395

Indexed by:

                 

                

                

  

 

The Prodia Education and Research Institute