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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
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Original Research Articles                      Volume : 6, Issue:2, February, 2017

PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
Publisher : Excellent Publishers
Email : editorijcmas@gmail.com /
submit@ijcmas.com
Editor-in-chief: Dr.M.Prakash
Index Copernicus ICV 2018: 95.39
NAAS RATING 2020: 5.38

Int.J.Curr.Microbiol.App.Sci.2017.6(2): 38-44
DOI: http://dx.doi.org/10.20546/ijcmas.2017.602.005


In-Vitro Culture and Characterization of Spermatogonial Stem Cells (SSCs) in Pre-Pubertal Mice
R. Kaavya1, T.A. Kannan2*, Sabiha Hayath Basha1, S. Vairamuthu3,Geetha Ramesh1 and B. Justin William2
1Department of Veterinary Anatomy, Tamil Nadu Veterinary and Animal Sciences University, Madras Veterinary College, Chennai- 600 007, India
2Centre for Stem Cell Research and Regenerative Medicine Tamil Nadu Veterinary and Animal Sciences University, Madras Veterinary College, Chennai- 600 007, India
3Centralised Instrumentation laboratory, Tamil Nadu Veterinary and Animal Sciences University, Madras Veterinary College, Chennai- 600 007, India
*Corresponding author
Abstract:

Spermatogonial Stem Cells (SSCs) are unipotent adult stem cells, defined as the undifferentiated stem cells located along the basement membrane of the seminiferous tubules. This present study is aimed to carry out the culture and characterization of SSCs in pre-pubertal age group of mice. Isolation of SSCs was done by two-step enzymatic digestion method and was seeded in a six-well plate. In primary culture (P0), colony forming unit was observed on day four post incubation. Mice Spermatogonial Stem Cells (mSSCs) adherence was observed on day eight post incubation. On twelve day post incubation, the cells were subcultured for further expansion. In the subsequent passages (P1 and P2) though a heterogenous morphology was observed, most of the cells showed spindle shaped structure. The pre-pubertal mSSCs were positive for alkaline phosphatase activity at P1 and P2 showed positive expression for the transcription factor, Oct4 was indicated by intranuclear green fluorescence. Flowcytometric analysis revealed that the mSSCs in the pre-pubertal age groups showed expression for the SSEA1 marker.


Keywords: Spermatogonial stem cells,Isolation, Culture, Characterization, Mice.

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How to cite this article:

Kaavya, R., T.A. Kannan, Sabiha Hayath Basha, S. Vairamuthu, Geetha Ramesh and Justin William, B. 2017. In-Vitro Culture and Characterization of Spermatogonial Stem Cells (SSCs) In Pre-Pubertal Mice.Int.J.Curr.Microbiol.App.Sci. 6(2): 38-44. doi: http://dx.doi.org/10.20546/ijcmas.2017.602.005
Copyright: This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license.

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