Vitrification of mouse oocytes using a nylon loop

Abstract

Cryopreservation of mouse oocytes was improved by the use of ultra‐rapid vitrification using a nylon loop of 0.5 mm diameter. Oocytes that were vitrified using the loop survived at high rates and were fertilized following a small hole being made in the zona pellucida (69.8%) and developed to the blastocyst stage in culture (67.4%) at similar rates to that of oocytes that were not cryopreserved. Blastocysts resulting from oocytes vitrified using the nylon loop had similar development of the inner cell mass and trophectoderm as blastocysts from non‐cryopreserved oocytes. In contrast, oocytes that were cryopreserved using a slow‐freezing protocol where most of the Na+ is replaced with choline had lower rates of fertilization (39.5%), reduced development to the blastocyst stage (25.7%), and blastocysts had reduced development of the inner cell mass. Blastocysts derived from oocytes that were vitrified with the nylon loop were able to implant (88.0%) and develop into fetuses (56.5%) at significantly higher rates compared to blastocysts derived from oocytes that were slow‐frozen (52.4 and 26.2%, respectively). Vitrification of mouse oocytes using the nylon loop results in the retention of viability of the oocytes and subsequent embryos. Mol. Reprod. Dev. 58:342–347, 2001. © 2001 Wiley‐Liss, Inc.

Cryopreservation of mammalian oocytes by using sugars: Intra- and extracellular raffinose with small amounts of dimethylsulfoxide yields high cryosurvival, fertilization, and development rates

“…An early study by Schroeder et al [63] represents an exception to this by demonstrating reasonably successful cryopreservation of mouse oocytes using a slow cooling protocol and 1.0 M DMSO. In the present study, the combination of intra- and extracellular raffinose with1/3 (0.5 M) and 2/3 (1.0M) of typical conventional CPA (Me 2 SO) concentration (i.e., 1.5 M) employed in slow cooling protocols yielded cryosurvival, fertilization and embryonic developmental rates that are similar to those of unfrozen controls and comparable to the best results in the literature achieved by using much higher cryoprotectant concentrations [7; 11; 41; 53; 69]. This indicates the outstanding efficiency of sugars along with small amounts of a conventional CPA in cryopreserving mammalian oocytes.…”

Cryopreservation of mammalian oocytes by using sugars: Intra- and extracellular raffinose with small amounts of dimethylsulfoxide yields high cryosurvival, fertilization, and development rates

“…An early study by Schroeder et al [63] represents an exception to this by demonstrating reasonably successful cryopreservation of mouse oocytes using a slow cooling protocol and 1.0 M DMSO. In the present study, the combination of intra- and extracellular raffinose with1/3 (0.5 M) and 2/3 (1.0M) of typical conventional CPA (Me 2 SO) concentration (i.e., 1.5 M) employed in slow cooling protocols yielded cryosurvival, fertilization and embryonic developmental rates that are similar to those of unfrozen controls and comparable to the best results in the literature achieved by using much higher cryoprotectant concentrations [7; 11; 41; 53; 69]. This indicates the outstanding efficiency of sugars along with small amounts of a conventional CPA in cryopreserving mammalian oocytes.…”

Vitrification within a nanoliter volume: oocyte and embryo cryopreservation within a 3D photopolymerized device

“…It is known that oocyte and embryo developmental potential is linked to their metabolism [ 46 ]. Previous studies have demonstrated that vitrification alters oocyte and embryo metabolism [ 47 , 48 ]. In these studies, metabolism was measured using metabolites present in spent media.…”

Tolerance of lamb and mouse oocytes to cryoprotectants during vitrification

“…It has been demonstrated that blastocyst development from vitrifiedwarmed buffalo oocytes fertilized by either IVF or ICSI showed similar embryo development (Liang et al, 2012;Attanasio et al, 2010). Conversely, Lane and Gardner (2001) reported that there was no significant difference in blastocyst development from vitrified-warmed oocytes when compared with unfrozen oocytes (96.3% vs 95.4% respectively). In this latter study, however, a different insemination method (zona drilling) was used for postwarmed oocytes, which possibly negating the zona hardening issue.…”

Cryopreservation of mammalian oocytes by using sugars: Intra- and extracellular raffinose with small amounts of dimethylsulfoxide yields high cryosurvival, fertilization, and development rates

“…An early study by Schroeder et al [63] represents an exception to this by demonstrating reasonably successful cryopreservation of mouse oocytes using a slow cooling protocol and 1.0 M DMSO. In the present study, the combination of intra- and extracellular raffinose with1/3 (0.5 M) and 2/3 (1.0M) of typical conventional CPA (Me 2 SO) concentration (i.e., 1.5 M) employed in slow cooling protocols yielded cryosurvival, fertilization and embryonic developmental rates that are similar to those of unfrozen controls and comparable to the best results in the literature achieved by using much higher cryoprotectant concentrations [7; 11; 41; 53; 69]. This indicates the outstanding efficiency of sugars along with small amounts of a conventional CPA in cryopreserving mammalian oocytes.…”

Vitrification within a nanoliter volume: oocyte and embryo cryopreservation within a 3D photopolymerized device

“…It is known that oocyte and embryo developmental potential is linked to their metabolism [ 46 ]. Previous studies have demonstrated that vitrification alters oocyte and embryo metabolism [ 47 , 48 ]. In these studies, metabolism was measured using metabolites present in spent media.…”

Tolerance of lamb and mouse oocytes to cryoprotectants during vitrification

“…It has been demonstrated that blastocyst development from vitrifiedwarmed buffalo oocytes fertilized by either IVF or ICSI showed similar embryo development (Liang et al, 2012;Attanasio et al, 2010). Conversely, Lane and Gardner (2001) reported that there was no significant difference in blastocyst development from vitrified-warmed oocytes when compared with unfrozen oocytes (96.3% vs 95.4% respectively). In this latter study, however, a different insemination method (zona drilling) was used for postwarmed oocytes, which possibly negating the zona hardening issue.…”

Cryopreservation of mammalian oocytes by using sugars: Intra- and extracellular raffinose with small amounts of dimethylsulfoxide yields high cryosurvival, fertilization, and development rates

“…An early study by Schroeder et al [63] represents an exception to this by demonstrating reasonably successful cryopreservation of mouse oocytes using a slow cooling protocol and 1.0 M DMSO. In the present study, the combination of intra- and extracellular raffinose with1/3 (0.5 M) and 2/3 (1.0M) of typical conventional CPA (Me 2 SO) concentration (i.e., 1.5 M) employed in slow cooling protocols yielded cryosurvival, fertilization and embryonic developmental rates that are similar to those of unfrozen controls and comparable to the best results in the literature achieved by using much higher cryoprotectant concentrations [7; 11; 41; 53; 69]. This indicates the outstanding efficiency of sugars along with small amounts of a conventional CPA in cryopreserving mammalian oocytes.…”

Vitrification within a nanoliter volume: oocyte and embryo cryopreservation within a 3D photopolymerized device

“…It is known that oocyte and embryo developmental potential is linked to their metabolism [ 46 ]. Previous studies have demonstrated that vitrification alters oocyte and embryo metabolism [ 47 , 48 ]. In these studies, metabolism was measured using metabolites present in spent media.…”

Tolerance of lamb and mouse oocytes to cryoprotectants during vitrification

“…It has been demonstrated that blastocyst development from vitrifiedwarmed buffalo oocytes fertilized by either IVF or ICSI showed similar embryo development (Liang et al, 2012;Attanasio et al, 2010). Conversely, Lane and Gardner (2001) reported that there was no significant difference in blastocyst development from vitrified-warmed oocytes when compared with unfrozen oocytes (96.3% vs 95.4% respectively). In this latter study, however, a different insemination method (zona drilling) was used for postwarmed oocytes, which possibly negating the zona hardening issue.…”