ArticleCryobiology2013
The effects of cooling rates and type of freezing extenders on cryosurvival of rat sperm.
Article in Cryobiology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
7 citing papers in PubMed, 34 citations in OpenAlex.
- A High-Resolution Microscopy System for Biological Studies of Cold-Adapted Species Under Physiological Conditions.Small methods · 2025Article
- Melatonin and selenium supplementation in extenders improves the post-thaw quality parameters of rat sperm.Clinical and experimental reproductive medicine · 2022Article
- Implementation of a method for sperm cryopreservation in sceloporine lizards.Conservation physiology · 2022Article
- Strategies for Highly Efficient Rabbit Sperm Cryopreservation.Animals : an open access journal from MDPI · 2021Review
- A new strategy and system for the ex vivo ovary perfusion and cryopreservation: An innovation.International journal of reproductive biomedicine · 2017Article
- Cryotolerance of Sperm from Transgenic Rhesus Macaques (Macaca mulatta).Journal of the American Association for Laboratory Animal Science : JAALAS · 2016Article
- Slow cryopreservation is not superior to vitrification in human spermatozoa; an experimental controlled study.Iranian journal of reproductive medicine · 2015Article
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
Abstract
Cryopreservation of rat sperm is very challenging due to its sensitivity to various stress factors. The objective of this study was to determine the optimal cooling rate and extender for epididymal sperm of outbred Sprague Dawley (SD) and inbred Fischer 344 (F344) rat strains. The epididymal sperm from 10 to 12 weeks old sexually mature SD and F344 strains were suspended in five different freezing extenders, namely HEPES buffered Tyrode's lactate (TL-HEPES), modified Kreb's Ringer bicarbonate (mKRB), 3% dehydrated skim milk (SM), Salamon's Tris-citrate (TRIS), and tes/tris (TES). All extenders contained 20% egg yolk, 0.75% Equex Paste and 0.1 M raffinose or 0.1 M sucrose. The sperm samples in each extender were cooled to 4°C and held for 45 min for equilibration before freezing. The equilibrated sperm samples in each extender were placed onto a shallow quartz dish inserted into Linkam Cryostage (BCS 196). The samples were then cooled to a final temperature of -150°C by using various cooling rates (10, 40, 70, and 100°C/min). For thawing, the quartz dish containing the sperm samples were rapidly removed from the Linkam cryo-stage and placed on a 37°C slide warmer and held for 1 min before motility analysis. Sperm membrane and acrosomal integrity and mitochondrial membrane potential (MMP) were assessed by SYBR-14/Propidium iodide, Alexa Fluor-488-PNA conjugate and JC-1, respectively. The total motility, acrosomal integrity, membrane integrity and MMP values were compared among cooling rates and extenders. Both cooling rate and type of extender had significant effect on cryosurvival (P < 0.05). Sperm motility increased as cooling rate was increased for both strains (P < 0.05). Highest cryosurvival was achieved when 100°C/min cooling rate was used in combination with TES extender containing 20% egg yolk, 0.75% Equex paste and either 0.1M sucrose or raffinose (P < 0.05). This study showed that TES extender containing 0.1 M raffinose or sucrose with 70°C/min and 100°C/min cooling rate improved post-thaw motility of rat sperm.
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Registered trials
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