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. 2009 Mar;26(2-3):143-9.
doi: 10.1007/s10815-009-9298-6. Epub 2009 Feb 7.

Achieving high survival rate following cryopreservation after isolation of prepubertal mouse spermatogonial cells

Firooz Jannat Alipoor  1 , Mohammad Ali Sadighi Gilani, Poopak Eftekhari-Yazdi, Ali Daliri Hampa, Hani Hosseinifar, Hiva Alipour, Mehdi Lotfi Panah
Affiliations

Achieving high survival rate following cryopreservation after isolation of prepubertal mouse spermatogonial cells

Firooz Jannat Alipoor et al. J Assist Reprod Genet. 2009 Mar.

Abstract

Purpose: Isolating spermatogonia cells with high purity and viability and achieving better survival rate following cryopreservation

Methods: Isolating the cells by Magnetic Activating Cell Sorting (MACS) method using anti CD49f (alpha6 integrin) antibody and Dynabeads and freezing in DMSO-based freezing mediums containing three different FBS concentrations of 50%, 60% and 70%.

Results: The mean (+/-SD) purity of the isolated cells was 92.52+/-3.57 (range 92.43-98.25). The cells frozen in group I, II and III had mean 39.60+/-1.48 (range 37.98-41.62), 89.05+/-3.83 (range 80.83-90.33) and 90.52+/-1.71 (range 89.07-92.52) viability, respectively.

Conclusion: Higher viable cell counts and purity can be attained by the use of alpha6 integrin and magnetic beads. After the thawing of spermatogonial cells, optimum viability was achieved in freezing media containing 60% FBS.

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Figures

Fig. 1
Fig. 1
a Seminiferous tubules after first step enzymatic digestion, magnification 100 ×s. b isolated cells after second enzymatic digestion. Magnification 400 ×s. c the cells after isolation by MACS method; these cells attached to magnetic beads and separated, magnification 1000×. d the cells that didn’t attach to magnetic beads and removed,magnification 1,000×
Fig. 2
Fig. 2
Cell purity analyzed by flow cytometry
See this image and copyright information in PMC

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References

    1. {'text': '', 'ref_index': 1, 'ids': [{'type': 'DOI', 'value': '10.1095/biolreprod.103.017012', 'is_inner': False, 'url': 'https://doi.org/10.1095/biolreprod.103.017012'}, {'type': 'PubMed', 'value': '12700182', 'is_inner': True, 'url': 'https://pubmed.ncbi.nlm.nih.gov/12700182/'}]}
    2. Kanatsu-Shinohara M, et al. Long-term proliferation in culture and germline transmission of mouse male germline stems cells. Biol Reprod. 2003;69(2):612–6. doi:10.1095/biolreprod.103.017012. - PubMed
    1. {'text': '', 'ref_index': 1, 'ids': [{'type': 'DOI', 'value': '10.1006/scdb.1998.0203', 'is_inner': False, 'url': 'https://doi.org/10.1006/scdb.1998.0203'}, {'type': 'PubMed', 'value': '9813187', 'is_inner': True, 'url': 'https://pubmed.ncbi.nlm.nih.gov/9813187/'}]}
    2. Griswold MD. The central role of Sertoli cells in spermatogenesis. Semin Cell Dev Biol. 1998;9(4):411–6. - PubMed
    1. {'text': '', 'ref_index': 1, 'ids': [{'type': 'DOI', 'value': '10.1093/humrep/dei412', 'is_inner': False, 'url': 'https://doi.org/10.1093/humrep/dei412'}, {'type': 'PubMed', 'value': '16311289', 'is_inner': True, 'url': 'https://pubmed.ncbi.nlm.nih.gov/16311289/'}]}
    2. Geens M, et al. Spermatogonial survival after grafting human testicular tissue to immunodeficient mice. Hum Reprod. 2006;21(2):390–6. doi:10.1093/humrep/dei412. - PubMed
    1. {'text': '', 'ref_index': 1, 'ids': [{'type': 'DOI', 'value': '10.1016/j.fertnstert.2003.07.021', 'is_inner': False, 'url': 'https://doi.org/10.1016/j.fertnstert.2003.07.021'}, {'type': 'PubMed', 'value': '14967371', 'is_inner': True, 'url': 'https://pubmed.ncbi.nlm.nih.gov/14967371/'}]}
    2. Agarwal A, et al. Fertility after cancer: a prospective review of assisted reproductive outcome with banked semen specimens. Fertil Steril. 2004;81(2):342–8. doi:10.1016/j.fertnstert.2003.07.021. - PubMed
    1. {'text': '', 'ref_index': 1, 'ids': [{'type': 'DOI', 'value': '10.1007/s003810050467', 'is_inner': False, 'url': 'https://doi.org/10.1007/s003810050467'}, {'type': 'PubMed', 'value': '10603019', 'is_inner': True, 'url': 'https://pubmed.ncbi.nlm.nih.gov/10603019/'}]}
    2. Bleyer WA. Epidemiologic impact of children with brain tumors. Childs Nerv Syst. 1999;15(11–12):758–63. doi:10.1007/s003810050467. - PubMed