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Abstracts: CryoLetters 42 (3), 2021

 

 

Volume 42 , No. 3 May/June 2021

ISSN 0143-2044

 

 


PERSPECTIVE: Cryopreservation of human oocytes and the 'carryover' effect on early embryo development (PDF)
Qi-Peng Jia and Wendell Q. Sun

120-128

 

 


Effects of cryopreservation on acrosin activity and DNA damage of Russian sturgeon (Acipenser gueldenstaedtii) semen
Xiaorong Huang, Tao Zhang, Feng Zhao, Guangpeng Feng,
Jianyi Liu, Gang Yang, Longzhen Zhang and Ping Zhuang

129-136

 

 


Status of beta defensin-1 and its effect on post thaw semen fertility gene expression in Indian goat breed
Ravi Ranjan, Pallavi Singh, Shiva Pratap Singh,
Kumaresan Gururaj, Suresh Dinkar Kharche and
Manoj Kumar Singh

137-145

 

 


Effect of idebenone, resveratrol and taurine on the sperm quality and lipid peroxidation of cryopreserved crossbred ram semen
Farooz Ahmad Lone, Mehrajuddin Naikoo,
Syed Mohmad Shah, Sajad Ahmad Darzi and Javid Farooq

146-153

 

 


Cryolipolysis with plate for treatment of localized adiposity
Veronica Bellocco, Daniela Rios, Daniela Podesta,
Rodrigo Marcel Valentim da Silva, Eneida de Morais Carreiro
and Patrícia Froes Meyer

154-158

 

 


Shoot tip cryopreservation as a conservation tool for species of Quercus: effects of species and environment on recovery
Valerie C. Pence and Mary F. Chaiken

159-167

 

 


Germplasm conservation techniques and assessment of physiological, biochemical and molecular integrity of indigenous, endangered Dioscorea prazeri
Smitha S. Thankappan and Villoo Morawala-Patell

168-177

 

 


Cryopreservation studies on silver carp (Hypophthalmichthys molitrix) embryos
Sheikh Mustafizur Rahman, Md. Ahsan Habib,
Abdur Rahman Khan, Md. Nazmul Ahsan,
Shaikh Tareq Arafat, Md. Moshiur Rahman,
Ahmed Saud Alsaqufi, Roshmon Thomas Mathew,
Yousof Naser Alrashada and Yousef Ahmed Alkhamis

178-187

 

 

 

 

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CryoLetters 42 (3), 120-128 (2020)
© CryoLetters,
editor@cryoletters.org

PERSPECTIVE:
CRYOPRESERVATION OF HUMAN OOCYTES AND THE 'CARRYOVER' EFFECT ON EARLY EMBRYO DEVELOPMENT 

Qi-Peng Jia and Wendell Q. Sun *

Institute of Biothermal Science and Technology, School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai, China.
Corresponding author's E-mail:
wendell.q.sun@gmail.com

Abstract

Worldwide women are increasingly facing the issue of delayed child-bearing and fertility decline. Oocyte cryopreservation provides an option for fertility preservation, especially for women with diseases and other special needs to conceive babies later. In this review we examine the effect of oocyte cryopreservation on early development of human embryos. Databases (Medline, PubMed and Web of Science) were searched for relevant clinical studies published between 1999 and 2020. A total of 27 studies on oocyte cryopreservation and embryo development were identified, and data in those studies are retrieved for meta-analysis on the outcomes of oocyte survival, fertilization and early embryo development. In comparison to the slow freezing technique, vitrification yields significantly better oocyte survival (84.7% ± 0.6% vs 58.0% ± 0.5%), and subsequently higher rates of fertilization (65.5% ± 0.9% vs 40.0% ± 0.6%), cleavage (58.8% ± 0.9% vs 34.6% ± 0.8%), as well as embryo implantation (5.9% ± 0.3% vs 2.9% ± 0.2%). This analysis reveals a negative 'carryover' effect of oocyte cryopreservation on early development of embryos after oocyte fertilization (i.e., cleavage and implantation). This 'carryover' effect is greater for slowly-frozen oocytes than for vitrified oocytes, and may represent subtle functional or molecular alterations that are not severe enough to affect cell survival and fertilization, but sufficient to impair later development. The nature of the 'carryover' effect is unknown. Hypothermia, membrane ion channels, bioenergy metabolism and epigenetic modifications are likely involved. In conclusion, oocyte cryopreservation can negatively affect early development of human embryos. Future studies should go beyond oocyte survival and look further into the effects on epigenetic changes.

Keywords: cryopreservation; embryo development; fertility preservation; oocyte; vitrification.

Download the paper: CRYOPRESERVATION OF HUMAN OOCYTES AND THE 'CARRYOVER' EFFECT ON EARLY EMBRYO DEVELOPMENT (PDF)

 

 

 

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CryoLetters 42 (3), 129-136 (2020)
© CryoLetters,
editor@cryoletters.org

EFFECTS OF CRYOPRESERVATION ON ACROSIN ACTIVITY AND DNA DAMAGE OF RUSSIAN STURGEON (ACIPENSER GUELDENSTAEDTII) SEMEN

Xiaorong Huang, Tao Zhang, Feng Zhao, Guangpeng Feng, Jianyi Liu,
Gang Yang, Longzhen Zhang and Ping Zhuang

Key Laboratory of East China Sea Fishery Resources Exploitation, Ministry of Agriculture, East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Shanghai, 200090, China; Shanghai Engineering Research Center of Fisheries Stock Enhancement and Habitat Restoration of the Yangtze Estuary, Shanghai, 2000900, China
*Corresponding author's E-mail:
pzhuang@ecsf.ac.cn

Abstract

BACKGROUND: Cryopreservation of sturgeon sperm can be successful, but there can be a decrease in sperm viability and the reasons are not clear. OBJECTIVE: To investigate variations in the acrosin activity and the DNA integrity of Acipenser gueldenstaedtii semen during cryopreservation at -196ºC. MATERIALS AND METHODS: Fish semen samples were randomly divided into three groups: [1] fresh control; [2] native semen diluted 1:1 with 23.4 mM sucrose + 0.25 mM KCl + 30 mM Tris (pH 8.0) and the addition of 10% methanol as cryoprotectant; and [3] semen without any diluents or cryoprotectants. Acrosin activity and DNA damage (COMET assay) were assessed. RESULTS: The average acrosin activity fell to 61% and 27% of the control for cryoprotected and non-cryoprotected semen after cryopreservation. The differences among the three groups were significant (P<0.05). We also observed that various indexes of DNA damage (L-tail; tail DNA, tail momentum, olive tail momentum) were higher in semen that had been frozen. CONCLUSION: Although cryopreservation of semen induces decreased acrosin activity and increased DNA damage, cryoprotectants can protect the semen during cryopreservation.

Keywords: Acipenser gueldenstaedtii; acrosin activity; cryopreservation; DNA damage; semen.

 

 

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CryoLetters 42 (3), 137-145 (2020)
© CryoLetters,
editor@cryoletters.org

STATUS OF BETA DEFENSIN-1 AND ITS EFFECT ON POST THAW SEMEN FERTILITY GENE EXPRESSION IN INDIAN GOAT BREED

Ravi Ranjan*, Pallavi Singh, Shiva Pratap Singh, Kumaresan Gururaj,
Suresh Dinkar Kharche and Manoj Kumar Singh

ICAR-Central Institute for Research on Goats, Makhdoom, Farah, Mathura, UP- 281122, India.
*Corresponding author's E-mail:
dr_raviranjan@yahoo.co.in; drraviranjan@gmail.com

Abstract

BACKGROUND: Defensins are antimicrobial peptides and uniformly spans the entire sperm surface and is not exclusive to a specific domain. Goat β-defensin-1 helps in initiation of motility and capacitation of sperm. OBJECTIVE: To know the status of β-defensin-1 in blood, semen and its effect on post thaw fertility gene expression in Indian goat breeds.  MATERIALS AND METHODS: Semen was extended and divided for estimation of β-defensin-1 and cryopreserved having different concentrations of β-defensin-1. RESULTS: Bet defensin-1 concentration (pg/mL) in neat semen, sperm pellet and seminal plasma was significantly higher (P<0.05) in goat breed Barbari followed by Jamunapari and Jakhrana. Beta defensing-1 was also high in Jakhrana blood followed by Barbari and Jamunapari. The post thaw motility, live sperm, acrosome intactness and hypo osmotic swelled sperms were significantly higher (P< 0.05) with 10 ng/mL β-defensin in the semen dilutor. CONCLUSION: Beta defensin (10 ng/mL) in semen dilutor may be used as immuno-modulator to get better post thaw quality suitable for artificial insemination.

Keywords: artificial insemination; beta defensin; gene expression; immunomodulatory; semen cryopreservation.

 

 

 

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CryoLetters 42 (3), 146-153 (2020)
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editor@cryoletters.org

EFFECT OF IDEBENONE, RESVERATROL AND TAURINE ON THE SPERM QUALITY AND LIPID PEROXIDATION OF CRYOPRESERVED CROSSBRED RAM SEMEN

Farooz Ahmad Lone1*, Mehrajuddin Naikoo1, Syed Mohmad Shah2,
Sajad Ahmad Darzi3 and Javid Farooq4

1Division of Animal Reproduction, Gynaecology and Obstetrics, Faculty of Veterinary Sciences and Animal Husbandry, Shuhama, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Srinagar-190006 UT Jammu & Kashmir, India.
2Krishi Vigyan Kendra, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Kargil-1, 194103, UT Ladakh, India.
3Frozen Semen Project, Ranbirbagh, Ganderbal- 191131 UT Jammu & Kashmir, India.
4Mountain Research Centre for Sheep & Goat, Faculty of Veterinary Sciences and Animal Husbandry, Shuhama, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Srinagar-190006 UT Jammu & Kashmir, India.
*Corresponding author's E-mail:
dr.farooz462@gmail.com

Abstract

BACKGROUND: Antioxidants reduce oxidative stress and improve sperm quality during cryopreservation. OBJECTIVE: To investigate the effect of idebenone, resveratrol and taurine on the sperm quality and lipid peroxidation of cryopreserved crossbred ram semen. METHODS: In a split study, pooled ejaculates were divided into four aliquots cryopreserved in tris extender with no antioxidant (control), with idebenone (0.01 mM), resveratrol (0.1 mM), and taurine (40 mM). RESULT: Among all antioxidants, taurine treatment yielded significantly better sperm quality. Malondialdehyde (MDA) level in seminal plasma was significantly lower for taurine compared to the control, idebenone and resveratrol treatments. Moreover, sperm quality declined significantly in all the groups from pre-freeze to post-thaw. CONCLUSION: The findings indicate that taurine at 40 mM significantly improves sperm quality compared to 0.01 mM idebenone and 0.1 mM resveratrol, hence it can be considered as a potent and promising antioxidant supplement in tris extender for the cryopreservation of crossbred ram semen.

Key words: antioxidants; cryopreservation; lipid peroxidation; ram; sperm quality.

 

 

 

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CryoLetters 42 (3), 154-158 (2020)
© CryoLetters,
editor@cryoletters.org

CRYOLIPOLYSIS WITH PLATE FOR TREATMENT OF LOCALIZED ADIPOSITY

Veronica Bellocco1, Daniela Rios2, Daniela Podesta3,
Rodrigo Marcel Valentim da Silva4*, Eneida de Morais Carreiro5
and Patrícia Froes Meyer5.

1 Córdoba Nacional University, Córdoba, Argentina,
2 Catholic University of Cuyo de San Juan, San Juan, Argentina,
3 Salvador's University, Buenos Aires, Argentina,
4 Federal University of Rio Grande do Norte, RN, Brasil
5 University Center of Rio Grande do Norte, Natal, Brasil.
*Corresponding author's E-mail:
rodrigomarcelvalentim@gmail.com

Abstract

BACKGROUND: Cryolipolysis is a non-invasive method capable of reducing the thickness of the fat layer. OBJECTIVE: To evaluate the effects of cryolipolysis with the use of plate applicators in the treatment of abdominal fat in women. MATERIALS AND METHODS: The sample was composed of 15 participants, who were evaluated before and at the end of the intervention. Three applications of cryolipolysis were performed in the infraumbilical portion of the abdominal region. The volunteers were divided into three groups G-1 (temperature of -2°C), G-2 (temperature of -3°C) and G3 (temperature of -4°C). RESULTS: There was a reduction in plicometry measurements in groups G2 and G3, in the comparison between the initial and final moments (p <0.05), and a reduction in perimetry and ultrasound (p <0.05) in all groups. It was found that the G3 group was subject to higher risk of first degree burns and redness when compared to the other groups. CONCLUSION: It is suggested that plate cryolipolysis is a possibly effective resource for reducing adiposity, as shown in the evaluation of perimetry, plicometry, and ultrasound results, and in the photographic analysis.

Keywords: cryolipolysis; localized adiposity; physiotherapy.

 

 

 

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CryoLetters 42 (3), 159-169 (2020)
© CryoLetters,
editor@cryoletters.org

SHOOT TIP CRYOPRESERVATION AS A CONSERVATION TOOL FOR SPECIES OF Quercus: EFFECTS OF SPECIES AND ENVIRONMENT ON RECOVERY

Valerie C. Pence* and Mary F. Chaiken

Center for Conservation and Research of Endangered Wildlife (CREW). Cincinnati Zoo and Botanical Garden, 3400 Vine Street Cincinnati, OH 45220 USA.
*Corresponding author's E-mail:
valerie.pence@cincinnatizoo.org

Abstract

BACKGROUND: The seeds of oak (Quercus) species cannot be conserved in conventional seed banks and require cryobiotechnologies for long-term, ex situ conservation. OBJECTIVE: To evaluate shoot tip cryopreservation as a method for conserving oaks ex situ. MATERIALS AND METHODS: Droplet vitrification was tested on tips from shoot cultures of four oak species; the effects of three preculture and recovery growth conditions on survival and growth were tested with Q. virginiana. RESULTS: Shoot tips of Q. virginiana gave the best survival through liquid nitrogen exposure (56%), compared with Q. hinckleyi (20%), Q. suber (12%), and Q. gambelii (0%). In further tests with Q. virginiana, recovery in the alternating temperature regime (27C/15C, 16 h / 8 h) gave significantly better survival than at constant 21C and significantly better growth than constant 26C. CONCLUSION: While there are species differences in the response to droplet vitrification, these results indicate that shoot tip cryopreservation should be explored further as a conservation tool for oaks.

Keywords: alternating temperature; droplet vitrification; exceptional species; in vitro; oak.

 

 

 

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CryoLetters 42 (3), 168-177 (2020)
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editor@cryoletters.org

GERMPLASM CONSERVATION TECHNIQUES AND ASSESSMENT OF PHYSIOLOGICAL, BIOCHEMICAL AND MOLECULAR INTEGRITY OF INDIGENOUS, ENDANGERED DIOSCOREA PRAZERI

Smitha S. Thankappan1* and Villoo Morawala-Patell2

1M S Swaminathan Research Foundation Community Agrobiodiversity Centre, Kalpetta, Wayanad, Kerala, India.
2Science, Innovation and Diagnostics division & Bio-Agriculture, Avesthagen Ltd, Bangalore, India, Affiliated to University of Mysore, Karnataka-570005, India.
*Corresponding author's E-mail:
smitha.st@gmail.com; smithastcabc@mssrf.res.in

Abstract

BACKGROUND: Cryopreservation is a reliable and economical method for the long-term ex situ conservation of valuable genetic resources. OBJECTIVE: The present study focuses on establishing novel regeneration strategies and on assessing various cryogenic methods using nodal explants/shoot apices and on developing in vitro technologies for germplasm conservation of Dioscorea prazeri. MATERIALS AND METHODS: Pre-treatment, growth regulators, temperature conditions, treatment period for recovery and growth of explants were optimized and various germplasm conservation methods were conducted to attain the conservation and mass multiplication of the endangered therapeutic plant. The plants regenerated from vitrified tissues were evaluated for physiological stability through morphological characteristics, genetic stability using RAPD analysis and with key metabolites for biochemical characterization. RESULTS: An optimized vitrification method resulted in a regeneration level of 92 ± 2 %, whereas a method comprising encapsulation dehydration resulted in 75 ± 2 % regeneration. In contrast, only a 38 ± 2 % regeneration was achieved using an encapsulation vitrification method. CONCLUSION: Vitrification-based procedures significantly improve cryopreservation survival and can be successfully employed for the long-term conservation of Dioscorea species and, potentially, other medicinal plants.

Keywords: diosgenin extraction; encapsulation dehydration; genetic fidelity assessment; HPLC analysis; nodal explants/shoot apices; vitrification.

 

 

 

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CryoLetters 42 (3), 178-187 (2020)
© CryoLetters,
editor@cryoletters.org

CRYOPRESERVATION STUDIES ON SILVER CARP (HYPOPHTHALMICHTHYS MOLITRIX) EMBRYOS

Sheikh Mustafizur Rahman1,2*, Md. Ahsan Habib1,
Abdur Rahman Khan1, Md. Nazmul Ahsan1, Shaikh Tareq Arafat1,
Md. Moshiur Rahman1, Ahmed Saud Alsaqufi3
Roshmon Thomas Mathew2, Yousof Naser Alrashada3
and Yousef Ahmed Alkhamis3

1Fisheries and Marine Resource Technology Discipline, Khulna University, Khulna-9208, Bangladesh
2Fish Resources Research Center, King Faisal University, Hofuf-420, Al-Ahsa, Kingdom of Saudi Arabia
3Department of Animal and Fish Production, College of Agricultural and Food Sciences, King Faisal University, Al-Ahsa, Kingdom of Saudi Arabia
*Corresponding author's E-mail:
mustafizfmrt@yahoo.com

Abstract

BACKGROUND: Cryopreservation is an effective tool for the preservation of live biological materials. OBJECTIVE: This study examined the suitability of cryopreservation protocols and the effectiveness of ultrasound for silver carp embryos. MATERIALS AND METHODS: Embryos at three developmental stages were exposed to 10, 15, 20, and 25% of five cryoprotectants (CPAs), namely propylene glycol (PG), dimethylformamide (DFA), DMSO, MeOH, and ethylene glycol (EG) for 20 min. Embryos were exposed to twelve vitrification solutions (VSs) for 10 (five steps of 2 min), 15 (five steps of 3 min), 20 (five steps of 4 min) min. Embryos were also exposed to ultrasound in VSs prior to cooling for cryopreservation. RESULTS: Hatching rates decreased with increasing CPA concentrations while toxicity varied in the order of PG < DMSO < EG < MeOH < DFA. Tail elongation stage was more tolerant to CPA than 6-somites and morula stages. The survival of embryos exposed to ultrasound in VS was remarkably lower than in water. Embryos exposed to ultrasound in VSs under the best conditions did not response well after attempted vitrification. CONCLUSION: Ultrasound-mediated CPA impregnation could be effective but other innovative methods may be needed to attain successful cryopreservation.

Keywords: embryo morphology; embryo toxicity; ultrasound; vitrification; vitrification solution.

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