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Article / Publication Details
Abstract
The present study aimed to identify the effects of sugar and methods [slow freezing (SF) vs fast freezing (FF)] on post-thaw in-vitro functional characteristics of cryopreserved caprine SSCs (cSSCs) and the cells obtained from cryopreserved testis tissue of pre-pubertal Barbari bucks. For this, in experiment-1, cSSCs were isolated and cryopreserved by either SF or FF method with different nonpermeable [sugars; trehalose (140 mM; 140T or 400 mM; 400T), and sucrose (140 mM; 140S or 400 mM; 400S)] or/and permeable [5% ethylene glycol (EG) and DMSO] cryoprotectants. After one-wk of cryopreservation, the cSSCs were thawed and cultured for evaluation of their characteristics. Further, in experiment-2, the effectiveness of sugars [trehalose (140 mM) or sucrose (140 mM)] for cryopreservation of testicular tissues of pre-pubertal Barbari bucks using the SF or FF method was evaluated. After one-wk of cryopreservation, the tissues were thawed and cSSCs were isolated and cultured for 3-wk. In both experiments, cSSCs were evaluated for recovery rate, proliferation, metabolic viability, senescence, and stemness markers expression. The recovery rate was 1.3, 1.3, and 1.1-folds higher in the 140T group compared with EG, 140S, and 400S groups, respectively. Similarly, the expression of stemness markers (PGP9.5 and OCT-4) was relatively higher in 140T group compared with the other groups. In experiment 2, the recovery rate of cells per unit tissue wt. was significantly (P
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Article / Publication Details
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