The effects of nonpenetrating cryoprotectants added to TEST-yolk-glycerol extender on the post-thaw motility of ram spermatozoa

Schmehl, Marcia K.; Vazquez, I.; Graham, E.F.

doi:10.1016/0011-2240(86)90060-x

Cited by 32 publications

(17 citation statements)

References 14 publications

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“…This approach would not seem to be very appropriate because it is well known that there are important differences in the physiological characteristics of epididymal vs ejaculated spermatozoa, specifically in their membrane properties, which affect the cell survival after cooling and freezing (Hammond, 1930; Walton, 1930; Watson and Morris, 1987; Rath and Niemann, 1997; Gilmore et al, 1998a; Lambrechts et al, 1999). Furthermore, epididymal spermatozoa have not been exposed to the complex secretions of the accessory sex glands; these secretions alter both the chilling sensitivity and the freezing resistance of ejaculated spermatozoa (Viruela and Rajaniemi, 1983; Schmehl et al, 1986). Consequently, the improvement of existing protocols for freezing and thawing deer epididymal spermatozoa is necessary.…”

mentioning

confidence: 99%

Effects of Thawing Procedure on Postthawed In Vitro Viability and In Vivo Fertility of Red Deer Epididymal Spermatozoa Cryopreserved at −196°C

Soler¹,

García

Fernández-Santos³

et al. 2003

Journal of Andrology

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ABSTRACT:In this study, we have determined the effects of individual factor and thawing procedure on in vitro viability and in vivo fertility of frozen-thawed red deer epididymal spermatozoa. The spermatozoa that were collected from 13 Iberian deer stags were diluted at room temperature in a Triladylா-20% egg yolk medium and frozen in nitrogen vapors. In the first experimental series, sperm samples were collected from 10 mature stags. For thawing, the frozen straws were subjected to 3 different procedures: I (37ЊC for 20 seconds), II (60ЊC for 8 seconds) and III (70ЊC for 5 seconds). Sperm cryosurvival was judged in vitro by microscopic assessments of individual sperm motility (SM) and of plasma membrane and acrosome (NAR) integrities. Statistically significant variations were found (P Ͻ .05) between stags for most of the seminal parameters evaluated. The thawing procedure did not have an effect on the seminal characteristics evaluated after this process, except for SM (P Ͻ .05), with the best overall recovery rates after freezing and thawing found with the use of protocol I. Our results also show a differential resistance to return to isosmotic conditions of spermatozoa thawed using the different thawing protocols. In the second experimental series (insemination artificial trial), with spermatozoa from 3 stags, results of fertility were statistically higher (69.7% vs 42.4%, P ϭ .014) when spermatozoa were thawed at 37ЊC for 20 seconds than were warmed at 60ЊC for 8 seconds. Therefore, thawing protocol I, which provides slow thawing rates, was the most beneficial for epididymal spermatozoa thawing of the cervid subspecies analyzed in this study. In summary, high in vitro survival and in vivo fertility of frozenthawed deer epididymal spermatozoa were dependent on warming rates, but each stag exhibited its own sensitivity to cryopreservation.

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confidence: 99%

Effects of Thawing Procedure on Postthawed In Vitro Viability and In Vivo Fertility of Red Deer Epididymal Spermatozoa Cryopreserved at −196°C

Soler¹,

García

Fernández-Santos³

et al. 2003

Journal of Andrology

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“…This problem can be circumvented by using reagents that do not penetrate the cell membrane, thus there is much interest in identifying the most efficacious non-membranepenetrating cryoprotectants. High molecular mass compounds have been tested for cryoprotecting potential for different types of cell (Ashwood-Smith et al, 1971;Schmehl et al, 1986;Sputtek et al, 1988;Phillips et al, 1996;Pellerin-Mendas et al, 1997;Singbartl et al, 1998;Henrich and Langer, 1999). Some of these compounds have the ability to protect the cell against freezing damage.…”

Section: Introductionmentioning

confidence: 99%

“…Dextran, a polymeric form of carbohydrate, has been reported as a cryoprotectant for the preservation of red blood cells (Ashwood-Smith et al, 1971;Pellerin-Mendas et al, 1997); however, few studies have been carried out on the preservation of sperm cells using dextran as cryoprotectant. In all these studies, dextran was used as a component of the complex extender (for example egg yolk) in the presence of glycerol, a well known cryoprotectant (Schmehl et al, 1986). Schmehl et al (1986) studied the effect of a small number of dextrans of different molecular masses (15-20, 70, 200-300 kDa) and hydrolysed dextran (0.8-1.6 kDa) using TEST-yolk-glycerol and ram semen (Schmehl et al 1986).…”

Section: Introductionmentioning

confidence: 99%

Effect of dextrans on cryopreservation of goat cauda epididymal spermatozoa using a chemically defined medium

Kundu¹

2002

Reproduction

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2000 kDa offered maximum cryorecovery of forward motility to the extent of approximately 23%, 21%, 19%, 18%, 16%, 15% and 8%, respectively. Optimum concentrations of these dextrans for cryoprotection of sperm motility were 8.42, 2.50, 1.09, 0.37, 0.31, 0.10 and 0.04 mmol l-1 , respectively. It thus appears that each dextran has a characteristic cryoprotection profile. The data show that the cryoprotecting efficacy and optimum cryoprotecting concentrations of dextrans are inversely related to their molecular mass. Dextran also served as a significant cryoprotectant in the presence of glycerol (0.87 mol l-1) and dimethyl sulphoxide (0.76 mol l-1), which are well known cryoprotectants; the action of the combined cryoprotectants was almost additive. The presence of glycerol or glycerol plus dimethyl sulphoxide caused a significant reduction (from 8.42 to 6.27 mmol l-1) in the optimum concentration of dextran. In the presence of the three cryoprotectants, recovery of sperm motility was as high as 58% (forward motility) and 60% (total motility).

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“…Dextran, a polymeric form of carbohydrate, has been reported as a cryoprotectant for the preservation of red blood cells (Ashwood-Smith et al, 1971;Pellerin-Mendas et al, 1997); however, few studies have been carried out on the preservation of sperm cells using dextran as cryoprotectant. In all these studies, dextran was used as a component of the complex extender (for example egg yolk) in the presence of glycerol, a well known cryoprotectant (Schmehl et al, 1986). Schmehl et al (1986) studied the effect of a small number of dextrans of different molecular masses (15)(16)(17)(18)(19)(20)70, and hydrolysed dextran (0.8-1.6 kDa) using TEST-yolk-glycerol and ram semen (Schmehl et al 1986).…”

mentioning

confidence: 99%

“…In all these studies, dextran was used as a component of the complex extender (for example egg yolk) in the presence of glycerol, a well known cryoprotectant (Schmehl et al, 1986). Schmehl et al (1986) studied the effect of a small number of dextrans of different molecular masses (15)(16)(17)(18)(19)(20)70, and hydrolysed dextran (0.8-1.6 kDa) using TEST-yolk-glycerol and ram semen (Schmehl et al 1986). The hydrolysed and 15-20 kDa dextran provided some cryoprotection (approximately 15-20%) for ram sperm cells (Schmehl et al 1986), but the other dextrans did not offer any significant cryoprotection.…”

mentioning

confidence: 99%

Effect of dextrans on cryopreservation of goat cauda epididymal spermatozoa using a chemically defined medium

Kundu¹,

Chakrabarty²,

Dutta³

et al. 2002

Reproduction

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Experiments were carried out to investigate the cryoprotecting potential of dextrans (ranging from 10 to 2000 kDa) using a synthetic model system developed recently in this laboratory. Goat spermatozoa from the cauda epididymidis were extracted in a chemically defined medium (modified Ringer's solution) and assayed for motility using a phase-contrast microscope. The sperm cells were subjected to a freezing protocol in a computer-controlled biofreezer (cooling at 0.25 degrees C min(-1) to 5 degrees C; 5 degrees C min(-1) to -20 degrees C; 20 degrees C min(-1) to -100 degrees C) and plunged into liquid nitrogen. The frozen cells were thawed rapidly at 37 degrees C in a thermostatic waterbath. In the absence of dextran, all the spermatozoa lost their motility. The cryoprotecting efficacy of each dextran was found to be biphasic in nature. Initially, as the concentration of dextran was increased, the recovery of sperm motility also increased and reached an optimum value; however, with further increases in dextran concentration, the recovery of motility decreased sharply. Of all the sugar polymers tested, 10 kDa dextran showed the highest cryoprotecting efficacy, whereas the 2000 kDa sugar polymer was the least active. Dextrans of 10, 40, 73, 173, 252, 500 and 2000 kDa offered maximum cryorecovery of forward motility to the extent of approximately 23%, 21%, 19%, 18%, 16%, 15% and 8%, respectively. Optimum concentrations of these dextrans for cryoprotection of sperm motility were 8.42, 2.50, 1.09, 0.37, 0.31, 0.10 and 0.04 mmol l(-1), respectively. It thus appears that each dextran has a characteristic cryoprotection profile. The data show that the cryoprotecting efficacy and optimum cryoprotecting concentrations of dextrans are inversely related to their molecular mass. Dextran also served as a significant cryoprotectant in the presence of glycerol (0.87 mol l(-1)) and dimethyl sulphoxide (0.76 mol l(-1)), which are well known cryoprotectants; the action of the combined cryoprotectants was almost additive. The presence of glycerol or glycerol plus dimethyl sulphoxide caused a significant reduction (from 8.42 to 6.27 mmol l(-1)) in the optimum concentration of dextran. In the presence of the three cryoprotectants, recovery of sperm motility was as high as 58% (forward motility) and 60% (total motility).

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The effects of nonpenetrating cryoprotectants added to TEST-yolk-glycerol extender on the post-thaw motility of ram spermatozoa

Cited by 32 publications

References 14 publications

Effects of Thawing Procedure on Postthawed In Vitro Viability and In Vivo Fertility of Red Deer Epididymal Spermatozoa Cryopreserved at −196°C

Effects of Thawing Procedure on Postthawed In Vitro Viability and In Vivo Fertility of Red Deer Epididymal Spermatozoa Cryopreserved at −196°C

Effect of dextrans on cryopreservation of goat cauda epididymal spermatozoa using a chemically defined medium

Effect of dextrans on cryopreservation of goat cauda epididymal spermatozoa using a chemically defined medium

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