ENRICHMENT OF UNDIFFERENTIATED TYPE A SPERMATOGONIA FROM GOAT TESTIS USING DISCONTINUOUS PERCOLL DENSITY GRADIENT AND DIFFERENTIAL PLATING

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HEIDARI BANAFSHEH; GIFANI MINOO; SHIRAZI ABOLFAZL; ZARNANI AMIR HASSAN; BARADARAN BEHZAD; NADERI MOHAMMAD MEHDI; BEHZADI BAHAREH; BORJIAN BOROUJENI SARA; SARVARI ALI; LAKPOUR NIKNAM; AKHONDI MOHAMMAD MEHDI

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Journal: Avicenna Journal of Medical Biotechnology Year:2014 | Volume:6 | Issue:2 Page(s): 94-103

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Title

ENRICHMENT OF UNDIFFERENTIATED TYPE A SPERMATOGONIA FROM GOAT TESTIS USING DISCONTINUOUS PERCOLL DENSITY GRADIENT AND DIFFERENTIAL PLATING

Author(s)

Keywords

C-KIT

DIFFERENTIAL PLATING

GOAT

PERCOLL GRADIENT

PGP 9.5

TYPE A SPERMATOGONIA

Abstract

Background: The well documented source for adult multipotent stem cells is Spermatogonial Stem Cells (SSCs). They are the foundation of spermatogenesis in the testis throughout adult life by balancing self-renewal and differentiation. The aim of this study was to assess the effect of percoll density gradient and DIFFERENTIAL PLATING on enrichment of undifferentiated TYPE A SPERMATOGONIA in dissociated cellular suspension of GOAT testes. Additionally, we evaluated the separated fractions of the gradients in percoll and samples in DIFFERENTIAL PLATING at different times for cell number, viability and purification rate of GOAT SSCs in culture. Methods: Testicular cells were successfully isolated from one month old GOAT testis using two-step enzymatic digestion and followed by two purification protocols, DIFFERENTIAL PLATING with different times of culture (3, 4, 5, and 6 hr) and discontinuous percoll density with different gradients (20, 28, 30, and 32%). The difference of percentage of undifferentiated SSCs (PGP9.5 positive) in each method was compared using ANOVA and comparison between the highest percentage of corresponding value between two methods was carried out by t-test using Sigma Stat (ver. 3.5).Results: The highest PGP9.5 (94.6±0.4) and the lowest C-KIT positive (25.1±0.7) in Percoll method was significantly (p≤0.001) achieved in 32% PERCOLL GRADIENT. While the corresponding rates in DIFFERENTIAL PLATING method for the highest PGP9.5 positive cells (81.3±1.1) and lowest C-KIT (17.1±1.4) was achieved after 5 hr culturing (p<0.001). The enrichment of undifferentiated type A spermato-gonia using Percoll was more efficient than DIFFERENTIAL PLATING method (p<0.001). Conclusion: Percoll density gradient and DIFFERENTIAL PLATING were efficient and fast methods for enrichment of TYPE A SPERMATOGONIAl stem cells from GOAT testes.

Cites

COMPARISON OF DIFFERENT ELECTROPORATION PARAMETERS ON TRANSFECTION EFFICIENCY OF SHEEP TESTICULAR CELLS

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APA: Copy

HEIDARI, BANAFSHEH, GIFANI, MINOO, SHIRAZI, ABOLFAZL, ZARNANI, AMIR HASSAN, BARADARAN, BEHZAD, NADERI, MOHAMMAD MEHDI, BEHZADI, BAHAREH, BORJIAN BOROUJENI, SARA, SARVARI, ALI, LAKPOUR, NIKNAM, & AKHONDI, MOHAMMAD MEHDI. (2014). ENRICHMENT OF UNDIFFERENTIATED TYPE A SPERMATOGONIA FROM GOAT TESTIS USING DISCONTINUOUS PERCOLL DENSITY GRADIENT AND DIFFERENTIAL PLATING. AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB), 6(2), 94-103. SID. https://sid.ir/paper/313836/en

Vancouver: Copy

HEIDARI BANAFSHEH, GIFANI MINOO, SHIRAZI ABOLFAZL, ZARNANI AMIR HASSAN, BARADARAN BEHZAD, NADERI MOHAMMAD MEHDI, BEHZADI BAHAREH, BORJIAN BOROUJENI SARA, SARVARI ALI, LAKPOUR NIKNAM, AKHONDI MOHAMMAD MEHDI. ENRICHMENT OF UNDIFFERENTIATED TYPE A SPERMATOGONIA FROM GOAT TESTIS USING DISCONTINUOUS PERCOLL DENSITY GRADIENT AND DIFFERENTIAL PLATING. AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB)[Internet]. 2014;6(2):94-103. Available from: https://sid.ir/paper/313836/en

IEEE: Copy

BANAFSHEH HEIDARI, MINOO GIFANI, ABOLFAZL SHIRAZI, AMIR HASSAN ZARNANI, BEHZAD BARADARAN, MOHAMMAD MEHDI NADERI, BAHAREH BEHZADI, SARA BORJIAN BOROUJENI, ALI SARVARI, NIKNAM LAKPOUR, and MOHAMMAD MEHDI AKHONDI, “ENRICHMENT OF UNDIFFERENTIATED TYPE A SPERMATOGONIA FROM GOAT TESTIS USING DISCONTINUOUS PERCOLL DENSITY GRADIENT AND DIFFERENTIAL PLATING,” AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB), vol. 6, no. 2, pp. 94–103, 2014, [Online]. Available: https://sid.ir/paper/313836/en

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