Volume 5, Issue 4 (7-2007)                   IJRM 2007, 5(4): 109-115 | Back to browse issues page

XML Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

Koruji M, Movahedin M, Mowla S J, Gourabi H. Colony formation ability of frozen thawed spermatogonial stem cell from adult mouse. IJRM 2007; 5 (4) :109-115
URL: http://ijrm.ir/article-1-81-en.html
1- Department of Anatomical Sciences, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2- Department of Anatomical Sciences, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran , mansoure@modares.ac.ir
3- Department of Genetics, School of Basic Sciences, Tarbiat Modares University, Tehran, Iran
4- Department of Genetics, Institute of Royan, Tehran, Iran
Abstract:   (2643 Views)
Background: The basis of spermatogenesis is the spermatogonial stem cells (SSCs). The concentration of SSCs is very small. However, a system that supports the proliferation and maintenance of SSCs in vitro could be used to preserve and expand SSCs numbers as well as increase success in transplantation. It is a new avenue to restore spermatogenesis in azoospermia subjects.
Objective: Proliferation and enhancement of frozen-thawed SSCs numbers during in vitro culture.
Materials and Methods: Both Sertoli and spermatogonial cells were isolated from adult mouse testes. Frozen-thawed spermatogonial cells were cultured in two groups: simple culture (Experimental 1) and co culture with Sertoli cells (Experimental 2). Also, Fresh cells were considered as control groups: simple culture (control1) and co culture with Sertoli cells (control 2).Assay of the spermatogonial-cell-derived colonies was carried out at the end of each week.
Results: Results indicated that the viability rate of the frozen cells after thawing (68.4±10.2%) was influenced by cryopreservation procedure significantly (p ≤0.001). In addition, the number of the colonies and their diameters in the co-culture system with fresh cells (25.1±5.2 and 205.8±50 µm, respectively) were more than other groups and the differences were significant (p<0.001). Number of the colonies and their diameters in experimental 1(9.5±4.3 and 124±35.9 µm, respectively), experimental 2 (15.6±3.5 and 157.6±41.9µm, respectively) groups were better than control 1 group (3.1±2.2 and 87.5±30.6µm, respectively) and the differences were significant (p<0.001).
Conclusion: We demonstrated that co-culture system with Sertoli cells can increase in vitro colony formation of adult fresh and frozen-thawed spermatogonial cells in mouse.
Full-Text [PDF 215 kb]   (518 Downloads) |   |   Full-Text (HTML)  (285 Views)  
Type of Study: Original Article |

1. Oatley JM, de Avila DM, Reeves JJ, McLean DJ. Testis tissue explant culture supports survival and proliferation of bovine spermatogonial stem cells. Biol Reprod 2004; 70: 625-631. [DOI:10.1095/biolreprod.103.022483]
2. Fuchs E, Segre JA. Stem cells: a new lease on life. Cell 2000; 100:143- 155. [DOI:10.1016/S0092-8674(00)81691-8]
3. Weissman IL. Translating stem and progenitor cell biology to the clinic: barriers and opportunities. Science 2000; 287:1442-1446. [DOI:10.1126/science.287.5457.1442]
4. Tegelenbosch RAJ, de Rooij DGA quantitative study of spermatogonial multiplication and stem cell renewal in the C3H: 101 F1 hybrid mouse. Mutat Res 1993; 290, 193-200. [DOI:10.1016/0027-5107(93)90159-D]
5. Nagano M, Brinster R.L. Spermatogonial transplantation and reconstitution of donor cell spermatogenesis in recipient mice, APMIS 1998; 106:47-55. [DOI:10.1111/j.1699-0463.1998.tb01318.x]
6. Izadyar F, Creemers LB, van Dissel-Emiliani FMF, de Rooij DG. Spermatogonial stem cell transplantation. Molecular and Cellular Endocrinology 2000; 169:21-25. [DOI:10.1016/S0303-7207(00)00346-4]
7. Maekawa M, Nishimune Y. In-vitro proliferation of germ cells and supporting cells in t he neonatal mouse testis. Cell Tissue Res 1991; 265: 551-554. [DOI:10.1007/BF00340879]
8. Anjamrooz SH, Movahedin M, Tiraihi T, Mowla SJ. In vitro effects of epidermal growth factor, follicle stimulating hormone and testosterone on mouse spermatogonial cell colony formation.Reproduction, Fertility and Development 2006;18, 709-720. [DOI:10.1071/RD05126]
9. Izadyar F, den Ouden K, Creemers LB, Posthuma G, Parvinen M, de Rooij DG. Proliferation and differentiation of bovine type A spermatogonia during long-term culture. Biol Reprod 2003; 68:272-281. [DOI:10.1095/biolreprod.102.004986]
10. van Pelt AMM, Morena AR, van Dissel-Emiliani FM, Boitani C, Gaemers IC, de Rooij DG, et al. Isolation of the synchronized A spermatogonia from adult vitamin A-deficient rat testes. Biol Reprod 1996; 55: 439-444. [DOI:10.1095/biolreprod55.2.439]
11. Scarpino S, Morena AR, Petersen C, Froysa B, Soder O, Boitani CA. Rapid method of Sertoli cells isolation by DSA lectin, allowing mitotic analyses. Mol Cell Endocrinol 1998; 146: 121-127. [DOI:10.1016/S0303-7207(98)00190-7]
12. Izadyar F, Matthijs-Rijsenbilt JJ, den Ouden K, Creemers LB, Woelders H, de Rooij DG. Development of a cryopreservation protocol for type A spermatogonia. J Androl 2002; 3:537-545.
13. Anway MD, Folmer J, Wright WW, Zirk BR. Isolation of Sertoli cells from Adult Rat Testes: An Approach to Ex Vivo Studies of Sertoli cells Function. Biol Reprod 2003; 68:996-1002 [DOI:10.1095/biolreprod.102.008045]
14. Kubota H, Avarbock MR, Brinster RL. Growth factors essential for self-renewal and expansion of mouse spermatogonial stem cells. Prroc Natl Accad Sci USA 2004; 101: 16489-16494. [DOI:10.1073/pnas.0407063101]
15. Palombi F, Dicarlo C. Alkaline Phosphatase is a Marker for Myoid Cells in Cultures of Rat Peritubular and Tubular Tissue. Biol Reprod 1988; 39: 1101-1109. [DOI:10.1095/biolreprod39.5.1101]
16. Jeong D, McLean DJ, Griswold MD. Long-term culture and transplantation of murine testicular germ cells. J Androl 2003; 24: 661-669. [DOI:10.1002/j.1939-4640.2003.tb02724.x]
17. Shi YQ, Wang QZ, Liao SY, Zhang Y, Liu YX, Han CS. In vitro propagation of spermatogonial stem cells from KM mice. Front Bio Sci 2006; 11, 2614-2622. [DOI:10.2741/1995]
18. Richards AJ, Enders GC, Resnick JL. Differentiation of murine premigratory primordial germ cells in culture. Biol Reprod 1999; 61: 1146-1151. [DOI:10.1095/biolreprod61.4.1146]
19. Schrans-Stassen BHGJ, van de Kant HJG, de Rooij DG, van Pelt AMM. Differential expression of c-kit in mouse undifferentiated and differentiating type A spermatogonia. Endocrinology 1999; 140:5894-5900. [DOI:10.1210/endo.140.12.7172]
20. Izadyar F, Spierenberg GT, Creemers LB, den Ouden K, de Rooij DG. Isolation and purificaton of type A spermatogonia from the bovine testis. Reprod 2002; 124:85-94. [DOI:10.1530/rep.0.1240085]
21. Mori C, Nakamura N, Dix DJ, Fujioka M, Nakagawa S, Shiota K, et al. Morphological analysis of germ cell apoptosis during postnatal testis development in normal and Hsp 70-2 knockout mice. Dev Dyn 1997; 208:125-136. https://doi.org/10.1002/(SICI)1097-0177(199701)208:1<125::AID-AJA12>3.0.CO;2-5 [DOI:10.1002/(SICI)1097-0177(199701)208:13.0.CO;2-5]
22. Oke BO, Suarez-Quian CA. Localization of secretory, membrane-associated and cytoskeletal proteins in rat testis using an improved immunocytochemical protocol that employs polyester wax. Biol Reprod 1993; 48: 621-631. [DOI:10.1095/biolreprod48.3.621]
23. Avarbock MR, Brinster CJ. Brinster RL. Reconstitution of spermatogenesis from frozen spermatogonial stem cells. Nat Med 1996; 2: 693-696. [DOI:10.1038/nm0696-693]
24. Brinster RL, Nagano M. Spermatogonial stem cell transplantation, cryopreservation and culture. Cell Dev Biol 1998; 9: 401-409. [DOI:10.1006/scdb.1998.0205]
25. Dobrinski I, Avarbock MR, Brinster RL. Germ cell transplantation from large domestic animals into mouse testes. Mol Reprod Dev 2000; 56: 270-279. https://doi.org/10.1002/1098-2795(200011)57:3<270::AID-MRD9>3.0.CO;2-Z [DOI:10.1002/1098-2795(200011)57:33.0.CO;2-Z]
26. Aponte PM, Soda T, van de Kant HJ, de Rooij DG. Basic features of bovine spermatogonial culture and effects of glial cell line-derived neurotrophic factor. Theriogenology 2006; 65: 1828-47. [DOI:10.1016/j.theriogenology.2005.10.020]
27. Creemers LB, den Ouden K, van Pelt AMM, de Rooij DG. Maintenance of adult mouse type A spermatogonia in vitro: influence of serum and growth factors and comparison with prepubertal spermatogonial cell culture. Reproduction 2002; 124: 791-99. [DOI:10.1530/rep.0.1240791]
28. Nagano M, Ryu BY, Brinster CJ, Avarbock MR, Brinster RL. Maintenance of mouse male germ line stem cells in vitro. Biol Reprod 2003; 68:2207-14. [DOI:10.1095/biolreprod.102.014050]

Send email to the article author

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Designed & Developed by : Yektaweb