Colonization of Mouse Spermatogonial Cells in Modified Soft Agar Culture System Utilizing Nanofibrous Scaffold: A New Approach

  • Ali Talebi Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran http://orcid.org/0000-0003-2067-6326
  • Mohammad Ali Sadighi Gilani Department of Urology, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran
  • Morteza Koruji Cellular and Molecular Research Center & Department of Anatomical Sciences, Iran University of Medical Sciences, Tehran, Iran
  • Jafar Ai Department of Tissue Engineering, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
  • Mohammad Jafar Rezaie Department of Embryology, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran
  • Shadan Navid Department of Anatomy, School of Medicine, Qom University of Medical Sciences, Qom, Iran
  • Majid Salehi Department of Tissue Engineering, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran
  • Mehdi Abbasi Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
DOI: https://doi.org/10.31661/gmj.v8i0.1319
Keywords: Adult Germline Stem Cells, Cell Proliferation, Tissue Scaffolds, Agar

Abstract

Background: Spermatogonial stem cells (SSCs) are considered in fertility management approaches of prepubertal boys facing cancer therapies. However, in vitro propagation has become an important issue due to a small number of SSCs in testicular tissue. The present study aimed to investigate a modified soft agar culture system by using a nanofibrous scaffold as a new approach to mimic in vivo conditions of SSCs development. Materials and Methods: The SSCs were isolated from neonate mouse testes, cultured on polycaprolactone scaffold, and covered by a layer of soft agar for 2 weeks. Then, the number and diameter of colonies formed in experimental groups were measured and spermatogonial markers (i.e., Plzf, Gfrα1, Id4, and c-Kit) in SSCs colonies were evaluated by a real-time polymerase chain reaction and immunostaining. Results: Our results indicated that the colonization rate of SSCs was significantly higher in the present modified soft agar culture system (P<0.05). Only Plzf indicated a significant increased at the levels (P<0.05), the gene expression levels of Id4, Plzf, and Gfrα1 were higher in the present culture system. In addition, the expression of the c-Kit gene as a differentiating spermatogonia marker was higher in presence of scaffold and soft agar compared with the amount of other experimental groups (P<0.05). Conclusion: The culture system by using nanofibrous scaffold and soft agar as a new culture method suggests the potential of this approach in SSCs enrichment and differentiation strategies for male infertility treatments, as well as in vitro spermatogenesis. [GMJ.2019;8:e1319]

References

Ward E, DeSantis C, Robbins A, Kohler B, Jemal A. Childhood and adolescent cancer statistics, 2014. CA Cancer J Clin. 2014;64(2):83-103.

https://doi.org/10.3322/caac.21219

PMid:24488779

Onofre J, Baert Y, Faes K, Goossens E. Cryopreservation of testicular tissue or testicular cell suspensions: a pivotal step in fertility preservation. Hum Reprod Update. 2016;22(6):744-61.

https://doi.org/10.1093/humupd/dmw029

PMid:27566839 PMCid:PMC5099994

Picton HM, Wyns C, Anderson RA, Goossens E, Jahnukainen K, Kliesch S et al. A European per-spective on testicular tissue cryopreservation for fertility preservation in prepubertal and adoles-cent boys. Hum Reprod. 2015;30(11):2463-75.

https://doi.org/10.1093/humrep/dev190

PMid:26358785

Galuppo AG. Spermatogonial stem cells as a therapeutic alternative for fertility preservation of prepubertal boys. Einstein (Sao Paulo). 2015;13(4):637-9.

https://doi.org/10.1590/S1679-45082015RB3456

PMid:26761559 PMCid:PMC4878644

He Y, Chen X, Zhu H, Wang D. Developments in techniques for the isolation, enrichment, main culture conditions and identification of spermatogonial stem cells. Cytotechnology. 2015;67(6):921-30.

https://doi.org/10.1007/s10616-015-9850-4

PMid:25749914 PMCid:PMC4628917

Boitani C, Di Persio S, Esposito V, Vicini E. Spermatogonial cells: mouse, monkey and man com-parison. Semin Cell Dev Biol. 2016;59:79-88.

https://doi.org/10.1016/j.semcdb.2016.03.002

PMid:26957475

Ibtisham F, Wu J, Xiao M, An L, Banker Z, Nawab A et al. Progress and future prospect of in vitro spermatogenesis. Oncotarget. 2017;8(39):66709-27.

https://doi.org/10.18632/oncotarget.19640

PMid:29029549 PMCid:PMC5630449

Khorshidi S, Solouk A, Mirzadeh H, Mazinani S, Lagaron JM, Sharifi S et al. A review of key chal-lenges of electrospun scaffolds for tissue-engineering applications. J Tissue Eng Regen Med. 2016;10(9):715-38.

https://doi.org/10.1002/term.1978

PMid:25619820

Dash TK, Konkimalla VB. Poly-є-caprolactone based formulations for drug delivery and tissue en-gineering: A review. J Controlled Release. 2012;158(1):15-33.

https://doi.org/10.1016/j.jconrel.2011.09.064

PMid:21963774

Mei X-X, Wang J, Wu J. Extrinsic and intrinsic factors controlling spermatogonial stem cell self-renewal and differentiation. Asian J Androl. 2015;17(3):347.

Stukenborg JB, Wistuba J, Luetjens CM, Elhija MA, Huleihel M, Lunenfeld E et al. Coculture of Spermatogonia With Somatic Cells in a Novel Three Dimensional Soft Agar Culture System. J An-drol. 2008;29(3):312-29.

https://doi.org/10.2164/jandrol.107.002857

PMid:18046051

Elhija MA, Lunenfeld E, Schlatt S, Huleihel M. Differentiation of murine male germ cells to sperma-tozoa in a soft agar culture system. Asian J Androl. 2011;14:285-93.

https://doi.org/10.1038/aja.2011.112

PMid:22057383 PMCid:PMC3735096

Navid S, Rastegar T, Baazm M, Alizadeh R, Talebi A, Gholami K et al. In vitro effects of melatonin on colonization of neonate mouse spermatogonial stem cells. Syst Biol Reprod Med. 2017;63(6):370-81.

https://doi.org/10.1080/19396368.2017.1358774

PMid:28846448

Salehi M, Naseri-Nosar M, Ebrahimi-Barough S, Nourani M, Khojasteh A, Hamidieh AA et al. Sciatic nerve regeneration by transplantation of Schwann cells via erythropoietin controlled-releasing pol-ylactic acid/multiwalled carbon nanotubes/gelatin nanofibrils neural guidance conduit. J Biomed Mater Res B Appl Biomater. 2017.

https://doi.org/10.1002/jbm.b.33952

PMid:28675568

Saeed F, Mahdi N-N, Hadi S, Simin M, Roksana T, Majid R et al. Taurine-loaded poly (ε-caprolactone)/gelatin electrospun mat as a potential wound dressing material: In vitro and in vivo evaluation. J Bioact Compatible Polym. 2017:0883911517737103.

Goossens E, Van Saen D, Tournaye H. Spermatogonial stem cell preservation and transplantation: from research to clinic. Hum Reprod. 2013;28(4):897-907.

https://doi.org/10.1093/humrep/det039

PMid:23427228

KarbalaeiMahdi A, Shahrousvand M, Javadi HR, Ghollasi M, Norouz F, Kamali M et al. Neural dif-ferentiation of human induced pluripotent stem cells on polycaprolactone/gelatin bi-electrospun nanofibers. Mater Sci Eng, C. 2017;78:1195-202.

https://doi.org/10.1016/j.msec.2017.04.083

PMid:28575957

Ghasemi-Mobarakeh L, Prabhakaran MP, Morshed M, Nasr-Esfahani M-H, Ramakrishna S. Elec-trospun poly (ɛ-caprolactone)/gelatin nanofibrous scaffolds for nerve tissue engineering. Bio-materials. 2008;29(34):4532-9.

https://doi.org/10.1016/j.biomaterials.2008.08.007

PMid:18757094

Chong E, Phan T, Lim I, Zhang Y, Bay B, Ramakrishna S et al. Evaluation of electrospun PCL/gelatin nanofibrous scaffold for wound healing and layered dermal reconstitution. Acta Bio-mater. 2007;3(3):321-30.

https://doi.org/10.1016/j.actbio.2007.01.002

PMid:17321811

Rahimi-Feyli P, Tajik P, Shafiei S, Dodel M, Arbabi F. The effect of poly L-lactic acid nanofiber on the induction of colony formation of frozen-thawed bovine spermatogonial stem cells in vitro. KAUMS Journal (FEYZ). 2015;19(1):15-23.

Eslahi N, Hadjighassem MR, Joghataei MT, Mirzapour T, Bakhtiyari M, Shakeri M et al. The effects of poly L-lactic acid nanofiber scaffold on mouse spermatogonial stem cell culture. Int J Nanomed-icine. 2013;8:4563-76.

https://doi.org/10.2147/IJN.S45535

PMid:24348035 PMCid:PMC3848747

Shakeri M, Kohram H, Shahverdi A, Shahneh AZ, Tavakolifar F, Pirouz M et al. Behavior of mouse spermatogonial stem-like cells on an electrospun nanofibrillar matrix. J Asst Rep Gen. 2013;30(3):325-32.

https://doi.org/10.1007/s10815-012-9916-6

PMid:23274510 PMCid:PMC3607679

Published
2019-05-09
How to Cite
Talebi, A., Sadighi Gilani, M. A., Koruji, M., Ai, J., Rezaie, M. J., Navid, S., Salehi, M., & Abbasi, M. (2019). Colonization of Mouse Spermatogonial Cells in Modified Soft Agar Culture System Utilizing Nanofibrous Scaffold: A New Approach. Galen Medical Journal, 8, e1319. https://doi.org/10.31661/gmj.v8i0.1319
Issue
Vol. 8 (2019): 2019
Section
Original Article

Authors who publish with this journal agree to the following terms:

  • Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution 4.0 International License that allows others to share the work with an acknowledgment of the work's authorship and initial publication in this journal.
  • Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in this journal.
  • Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).