Volume 7, Issue 28 And 29 (Autumn & Winter 2009)                   ASJ 2009, 7(28 And 29): 113-120 | Back to browse issues page

XML Print


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

Janat Alipour F, Sedighi Gilani M A, Eftekhari Yazdi P, Daneshzadeh M T, Khakzad H. The Effect of FBS Concentration on Cryopreservation of Isolated Spermatogonial Cells from Neonatal Mouse by Macs Method. ASJ 2009; 7 (28 and 29) :113-120
URL: http://anatomyjournal.ir/article-1-494-en.html
Abstract:   (433 Views)
Purpose: The purpose of this study was the separation of spermatogonial cells from prepubertal NMRI mouse testis by magnetic activating cell sorting (MACS) method and assessment of FBS concentration on surviving these cells after cryopreservation. Materials and Methods: Testes from 6-days-old mouse removed and digested in two enzymatic mediums. First 8-9 min at medium containing collagenase, trypsin and DNase and second 10-12 min in medium containing collagenase, trypsin, DNase, hyalloronidase and EDTA and then the spermatogonial cells were isolated by (MACS) method. In the freezing step, the cells were frozen in three medium containing DMEM/F12 medium, 10% DMSO and 50%, 60% and 70% FBS serum, named group I, II. III.Results: The viable cells obtained after enzymatic disaggregation were 91.66%±0.60% and after being isolated by MACS method were 95.25%±0.33%. The purity of the isolated cells was 93.79%±2.20%. The cells frozen in group I, II and III had 39.09%±0.15, 85.55%±6.98 and 90.29±1.38 % viability, respectively.Conclusion: according to the obtained results, the increase of temperature at digestion step reduces the time of testicular tissue disaggregation and consequently increases viable cells. Higher viable cells and purity can be attained by using of α6 integrin and magnetic beads. The results show that spermatogonial cells in freezing media containing 60%-70% FBS have the highest viability after thawing.
Full-Text [PDF 333 kb]   (126 Downloads)    
Type of Study: Original | Subject: Morphometry
Received: 2021/12/27 | Accepted: 2009/11/28 | Published: 2009/11/28

Add your comments about this article : Your username or Email:
CAPTCHA

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

Designed & Developed by : Yektaweb