845 results match your criteria slow-freezing cryopreservation

Delivering embryos following 10 years of cryopreservation, using unpaired freeze/thaw techniques: A case report.

JBRA Assist Reprod 2021 Jun 9. Epub 2021 Jun 9.

Monteleone Centro de Reprodução Humana, São Paulo, SP, Brazil.

Although frozen embryo transfer is a widely established route for assisted reproduction, successful frozen embryo transfer using embryos that have undergone long term cryopreservation remains relatively unexplored, and its efficacy remains a matter of some debate. This case report describes two successful frozen embryo transfer conceptions in the same patient, one after 3 months of cryopreservation and the second 10 years after cryopreservation. These embryos were cryopreserved using the slow freezing technique and were thawed using an unpaired technique (ultra-rapid warming) after 10 years of storage. Read More

View Article and Full-Text PDF

Red Blood Cell Cryopreservation with Minimal Post-Thaw Lysis Enabled by a Synergistic Combination of a Cryoprotecting Polyampholyte with DMSO/Trehalose.

Biomacromolecules 2021 Jun 7. Epub 2021 Jun 7.

Department of Chemistry, University of Warwick, Coventry CV4 7AL, U.K.

From trauma wards to chemotherapy, red blood cells are essential in modern medicine. Current methods to bank red blood cells typically use glycerol (40 wt %) as a cryoprotective agent. Although highly effective, the deglycerolization process, post-thaw, is time-consuming and results in some loss of red blood cells during the washing procedures. Read More

View Article and Full-Text PDF

Lipid profile of bovine grade-1 blastocysts produced either in vivo or in vitro before and after slow freezing process.

Sci Rep 2021 Jun 2;11(1):11618. Epub 2021 Jun 2.

CNRS, IFCE, INRAE, Université de Tours, PRC, 37380, Nouzilly, France.

Currently, in vitro embryo production (IVP) is successfully commercially applied in cattle. However, the high sensitivity of embryos to cryopreservation in comparison to in vivo (IVD) embryos slows the dissemination of this biotechnology. Reduced cryotolerance is frequently associated with lipid accumulation in the cytoplasm mainly due to in vitro culture conditions. Read More

View Article and Full-Text PDF

Synergistic effect of Royal jelly in combination with glycerol and dimethyl sulfoxide on cryoprotection of Romanov ram sperm.

Cryobiology 2021 May 30. Epub 2021 May 30.

Biotechnology Laboratory, National Breeding Center and Improvement of Animal Production, P.O.Box 31585-963, Meshkindasht Road, Karaj, Iran.

Sperm fertility decreases significantly after freezing. Providing a suitable and useful diluent compound for freezing ram sperm can increase the efficiency of artificial insemination and consequently, the reproductive performance of sheep. Various biological properties such as antibacterial, anti-cancer, immunosuppressive, antioxidant and reproductive properties of royal jelly (RJ) are well known. Read More

View Article and Full-Text PDF

In Situ Vitrification of Lung Cancer Organoids on a Microwell Array.

Micromachines (Basel) 2021 May 28;12(6). Epub 2021 May 28.

Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Biological Science and Medical Engineering, Beihang University, Beijing 100083, China.

Three-dimensional cultured patient-derived cancer organoids (PDOs) represent a powerful tool for anti-cancer drug development due to their similarity to the in vivo tumor tissues. However, the culture and manipulation of PDOs is more difficult than 2D cultured cell lines due to the presence of the culture matrix and the 3D feature of the organoids. In our other study, we established a method for lung cancer organoid (LCO)-based drug sensitivity tests on the superhydrophobic microwell array chip (SMAR-chip). Read More

View Article and Full-Text PDF

Cryopreservation and Preparation of Thawed Spermatozoa from Rhesus Macaques (Macaca mulatta) for In Vitro Fertilization.

J Am Assoc Lab Anim Sci 2021 May 23. Epub 2021 May 23.

Advances in assisted reproductive technologies in rhesus macaques have allowed the development of valuable models of human disease, particularly when combined with recent techniques for gene editing. While the ability to perform in vitro fertilization (IVF) in rhesus macaques is well established, this procedure has not yet been optimized. Specifically, damage to the sperm caused by cryopreservation (cryodamage) may lead to unsuccessful artificial insemination and low fertilization and blastocyst formation rates in vitro. Read More

View Article and Full-Text PDF

Influence of hydrogel encapsulation during cryopreservation of ovarian tissues and impact of post-thawing in vitro culture systems in a research animal model.

Clin Exp Reprod Med 2021 Jun 24;48(2):111-123. Epub 2021 May 24.

Division of Reproductive Medicine, Department of Obstetrics and Gynecology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.

Objective: Using domestic cats as a biomedical research model for fertility preservation, the present study aimed to characterize the influences of ovarian tissue encapsulation in biodegradable hydrogel matrix (fibrinogen/thrombin) on resilience to cryopreservation, and static versus non-static culture systems following ovarian tissue encapsulation and cryopreservation on follicle quality.

Methods: In experiment I, ovarian tissues (n=21 animals; 567 ovarian fragments) were assigned to controls or hydrogel encapsulation with 5 or 10 mg/mL fibrinogen (5 or 10 FG). Following cryopreservation (slow freezing or vitrification), follicle viability, morphology, density, and key protein phosphorylation were assessed. Read More

View Article and Full-Text PDF

Vitrification yields higher cryo-survival rate than slow freezing in biopsied bovine in vitro produced blastocysts.

Theriogenology 2021 Sep 13;171:44-54. Epub 2021 May 13.

Department of Veterinary and Animal Sciences, University of Copenhagen, 1870 Frederiksberg C, Denmark. Electronic address:

Vitrification and slow freezing are the two commonly used embryo cryopreservation methods. In most studies, vitrification of intact embryos has proven superior in several respects, including cell and embryo survival and pregnancy rate. However, there is a lack of data for comparing these two methods in in vitro produced (IVP) bovine blastocysts, which have been subjected to the retrieval of trophectoderm (TE) biopsy. Read More

View Article and Full-Text PDF
September 2021

Comparison of two methods for prolong storage of decellularized mouse whole testis for tissue engineering application: An experimental study.

Int J Reprod Biomed 2021 Apr 22;19(4):321-332. Epub 2021 Apr 22.

Basic Medical Science Research Center, Histogenotech Company, Tehran, Iran.

Background: Biological scaffolds are derived by the decellularization of tissues or organs. Various biological scaffolds, such as scaffolds for the liver, lung, esophagus, dermis, and human testicles, have been produced. Their application in tissue engineering has created the need for cryopreservation processes to store these scaffolds. Read More

View Article and Full-Text PDF

Gene Expression of Fresh and Frozen/Thawed Canine Embryos.

Cryo Letters 2020 Nov-Dec;41(6):330-336

Department of Radiology and Animal Reproduction, FMVZ/UNESP, Botucatu/SP, Brazil.

Background: Canine embryo cryopreservation and subsequent transfer are relevant in the use of reproductive technologies.

Objective: The purpose of this study is the identification and quantification of the gene expression BAX and Bcl2, AQP3, Na+/K+ ATPase alpha-1 and beta-1 and LIFr in canine embryos obtained in vivo and after freezing.

Materials And Methods: For the collection of embryos, the bitches were identified at pro-estrous until the detection of 80-90% superficial cells. Read More

View Article and Full-Text PDF

Survivable potential of germ cells after trehalose cryopreservation of bovine testicular tissues.

Cryobiology 2021 May 11. Epub 2021 May 11.

College of Veterinary Medicine, Jilin University, Changchun, 130062, Jilin, China. Electronic address:

Germplasm preservation of livestock or endangered animals and expansion of germline stem cells are important. The purpose of this study is to investigate whether supplementation of trehalose to the freezing medium (FM) reduces tissular damage and improves the quality of testicular cells in the cryopreserved bovine testicular tissues. We herein established an optimized protocol for the cryopreservation of bovine testicular tissues, and the isolation as well as culture of bovine germ cells containing spermatogonial stem cells (SSCs) from these tissues. Read More

View Article and Full-Text PDF

PERSPECTIVE: Cryopreservation of Human Oocytes and the 'Carryover' Effect on Early Embryo Development.

Q P Jia W Q Sun

Cryo Letters 2021 May-Jun;42(3):120-128

Institute of Biothermal Science and Technology, School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai, China.

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. Read More

View Article and Full-Text PDF

Evaluation of quality and gene expression of goat embryos produced in vivo and in vitro after cryopreservation.

Cryobiology 2021 May 5. Epub 2021 May 5.

Laboratory of Reproductive Biotechniques, Department of Veterinary Medicine, Federal Rural University of Pernambuco, Brazil. Electronic address:

In the present study, we aimed to identify morphological and molecular changes of in vivo and in vitro-produced goat embryos submitted to cryopreservation. In vivo embryos were recovered by transcervical technique from superovulated goats, whereas in vitro produced embryos were produced from ovaries collected at a slaughterhouse. Embryos were frozen by two-steps slow freezing method, which is defined as freezing to -32 °C followed by transfer to liquid nitrogen. Read More

View Article and Full-Text PDF

A Tool for Accurate Stoichiometric Composition of Cryopreservative Media for Fetal and Induced Pluripotent Stem Cell-Derived Human Neural Stem Cells.

Curr Protoc 2021 May;1(5):e123

Center for Stem Cells and Regenerative Medicine, Sanford Burnham Prebys Medical Discovery Institute, Sanford Consortium for Regenerative Medicine, La Jolla, California.

Fetal human neural stem cells (fhNSC) are of considerable interest as potential regenerative therapies for neuronal or glial degeneration or destruction resulting from genetic abnormalities, disease, or injury. Realization of this potential requires securing a supply of cells sufficient to meet the needs of transplantation, which are often tens to hundreds of millions of cells per dose. This challenge necessitates the establishment of safe and efficient cell banking protocols. Read More

View Article and Full-Text PDF

Methods of Ovarian Tissue Cryopreservation: Is Vitrification Superior to Slow Freezing?-Ovarian Tissue Freezing Methods.

Reprod Sci 2021 May 3. Epub 2021 May 3.

Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, University of Florida, Gainesville, FL, USA.

After cancer treatment, female survivors often develop ovarian insufficiency or failure. Oocyte and embryo freezing are well-established fertility preservation options, but cannot be applied in pre-pubescent girls, in women with hormone-sensitive malignancies, or when gonadotoxic treatment cannot be delayed. Although ovarian tissue cryopreservation (OTC) has been used to restore fertility and endocrine function, the relative efficacy of its two major protocols, slow freezing and vitrification, remains controversial. Read More

View Article and Full-Text PDF

Influence of freezing techniques and glycerol-based cryoprotectant combinations on the survival of testicular tissues from adult collared peccaries.

Theriogenology 2021 Jun 26;167:111-119. Epub 2021 Mar 26.

Laboratory of Animal Germplasm Conservation, Federal Rural University of Semi-Arid - UFERSA, Mossoró, RN, Brazil. Electronic address:

The objective of the study was to evaluate the effects of different cryopreservation techniques including glycerol-based cryoprotectant combinations on the structure and viability of testicular tissues from adult collared peccaries. Tissue biopsies (3.0 mm³) from 5 different individuals were allocated to 10 different groups: fresh control; slow freezing (SF), conventional vitrification (CV), or solid-surface vitrification (SSV); each of them using three different combinations of cryoprotectants [dimethyl sulfoxide (DMSO) + ethylene glycol (EG); DMSO + Glycerol; and EG + Glycerol]. Read More

View Article and Full-Text PDF

Coaxial Alginate Hydrogels: From Self-Assembled 3D Cellular Constructs to Long-Term Storage.

Int J Mol Sci 2021 Mar 18;22(6). Epub 2021 Mar 18.

Institute for Multiphase Processes, Leibniz University Hannover, An der Universität 1, Building 8143, 30823 Garbsen, Germany.

Alginate as a versatile naturally occurring biomaterial has found widespread use in the biomedical field due to its unique features such as biocompatibility and biodegradability. The ability of its semipermeable hydrogels to provide a favourable microenvironment for clinically relevant cells made alginate encapsulation a leading technology for immunoisolation, 3D culture, cryopreservation as well as cell and drug delivery. The aim of this work is the evaluation of structural properties and swelling behaviour of the core-shell capsules for the encapsulation of multipotent stromal cells (MSCs), their 3D culture and cryopreservation using slow freezing. Read More

View Article and Full-Text PDF

The Effectiveness of Anti-Apoptotic Agents to Preserve Primordial Follicles and Prevent Tissue Damage during Ovarian Tissue Cryopreservation and Xenotransplantation.

Int J Mol Sci 2021 Mar 3;22(5). Epub 2021 Mar 3.

Department of Obstetrics and Gynecology, Korea University College of Medicine 73, Inchon-ro, Seongbuk-gu, Seoul 02841, Korea.

The purpose of this study is to investigate the effectiveness of sphingosine-1-phosphate (S1P) and Z-VAD-FMK (Z-VAD) as anti-apoptotic agents to preserve ovarian function and prevent tissue damage during ovarian tissue cryopreservation and transplantation. This study consisted of two steps, in vitro and in vivo. In the first step, human ovarian tissues were cryopreserved using slow-freezing media alone, S1P, or Z-VAD (control, S1P, Z-VAD group); based on the outcomes in these groups, Z-VAD was selected for subsequent xenotransplantation. Read More

View Article and Full-Text PDF

Influence of circulating testosterone concentration on sperm cryoresistance: The ibex as an experimental model.

Andrology 2021 Mar 9. Epub 2021 Mar 9.

Department of Animal Reproduction, INIA, Madrid, Spain.

Background: Recent studies have noted that the circulating testosterone concentration may affect the ability of spermatozoa to survive cryopreservation. However, few attempts to confirm such a relationship have been made. Wild ruminant species have very marked seasonal changes in their reproductive function and strong annual changes in their plasma testosterone concentration. Read More

View Article and Full-Text PDF

A Method to Efficiently Cryopreserve Mammalian Cells on Paper Platforms.

Bio Protoc 2020 Sep 20;10(18):e3764. Epub 2020 Sep 20.

Division of Engineering, New York University Abu Dhabi, Abu Dhabi, UAE.

This protocol describes a simple method to cryopreserve mammalian cells within filter papers as an alternative to conventional slow-freezing approach. The method involves treating paper fibers with fibronectin, using low concentrations of the cryoprotectant dimethyl sulfoxide (DMSO), and slow freezing cells to -80 °C at a 1 °C min rate. In our method, the biocompatibility, large surface area, 3D porosity and fiber flexibility of the paper, in combination with the fibronectin treatment, yield recovery of cells comparable to conventional approaches, with no additional fine-tuning to freezing and thawing procedures. Read More

View Article and Full-Text PDF
September 2020

Cryopreservation of Porcine Embryos: Recent Updates and Progress.

Biopreserv Biobank 2021 Jun 24;19(3):210-218. Epub 2021 Feb 24.

National Engineering Laboratory for Animal Breeding, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing, China.

Cryopreservation of embryos is important for long-distance embryo transfer and conservation of genetic resources. Porcine research is important for animal husbandry and biomedical research. However, porcine embryos are difficult to cryopreserve because of their high cytoplasmic lipid content and sensitivity to chilling stress. Read More

View Article and Full-Text PDF

Effects of slow freezing and vitrification on embryo development in domestic cat.

Reprod Domest Anim 2020 Oct 11;55(10):1328-1336. Epub 2020 Aug 11.

Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia.

Cryopreservation of gametes and embryos is used to maintain genetic diversity of domestic and wild felids. However, felid oocytes and preimplantation embryos contain large amount of intracellular lipids, which affect their cryosensitivity. The objective was to compare the effects of slow freezing and vitrification and to study lipid phase transition (LPT) during cooling in cat embryos. Read More

View Article and Full-Text PDF
October 2020

Elective oocyte cryopreservation for age-related fertility decline.

J Assist Reprod Genet 2021 May 19;38(5):1177-1186. Epub 2021 Feb 19.

Homerton Fertility Centre, Homerton University Hospital, Homerton Row, Clapton, London, E9 6SR, UK.

Purpose: Women who pursue fertility at an advanced age are increasingly common. Family planning and sexual education have traditionally focused on contraception and prevention of sexually transmitted diseases. A focus should now also be placed on fertility awareness and fertility preservation. Read More

View Article and Full-Text PDF

Diverse Approaches to Ovarian Tissue Cryopreservation Have Equivalent Outcomes in Markers of Tissue Viability.

Reprod Sci 2021 Feb 18. Epub 2021 Feb 18.

Women's Health Institute, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, OH, 44915, USA.

Ovarian tissue cryopreservation (OTC) is an accepted method of fertility preservation. However, OTC is not standardized and many variations exist in the freezing strategy, tissue processing, and surgical approach. In this pilot study, we used a sheep model to compare slow freezing versus vitrification techniques, as well as the feasibility of processing ovarian tissue into a hyaluronan suspension of small ovarian units. Read More

View Article and Full-Text PDF
February 2021

Cryopreservation of plant cell cultures - Diverse practices and protocols.

N Biotechnol 2021 May 14;62:86-95. Epub 2021 Feb 14.

Department Bioprocess Engineering, Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Forckenbeckstrasse 6, 52074, Aachen, Germany; Institute for Molecular Biotechnology, RWTH Aachen University, Worringerweg 1, 52074, Aachen, Germany. Electronic address:

Plant cell cultures can be used as biotechnological platforms for the commercial production of small-molecule active ingredients and recombinant proteins, such as biopharmaceuticals. This requires the cryopreservation of well-characterized cell lines as master cell banks from which uniform working cell banks can be derived to ensure high batch-to-batch reproducibility during production campaigns. However, the cryopreservation of plant cells is challenging due to their low viability and poor regrowth after thawing. Read More

View Article and Full-Text PDF

Ovarian tissue cryopreservation and novel bioengineering approaches for fertility preservation.

Curr Breast Cancer Rep 2020 Dec 4;12(4):351-360. Epub 2020 Nov 4.

Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan, United States.

Purpose Of Review: Breast cancer patients who cannot delay treatment or for whom hormone stimulation and egg retrieval are contraindicated require alternative methods of fertility preservation prior to gonadotoxic treatment. Ovarian tissue cryopreservation is an alternative approach that may offer patients the opportunity to preserve fertility and carry biologically-related children later in life. Various experimental approaches are being explored to obtain mature gametes from cryopreserved and thawed ovarian tissue for fertilization and implantation using biomimetic tissue culture Here we review the most recent developments in ovarian tissue cryopreservation and exciting advances in bioengineering approaches to tissue and ovarian follicle culture. Read More

View Article and Full-Text PDF
December 2020

Improved cryopreservation of in vitro produced bovine embryos using FGF2, LIF, and IGF1.

PLoS One 2021 3;16(2):e0243727. Epub 2021 Feb 3.

Division of Animal Sciences, University of Missouri, Columbia, Missouri, United States of America.

In vitro embryo production systems are limited by their inability to consistently produce embryos with the competency to develop to the blastocyst stage, survive cryopreservation, and establish a pregnancy. Previous work identified a combination of three cytokines [fibroblast growth factor 2 (FGF2), leukemia inhibitory factor (LIF), and insulin-like growth factor 1 (IGF1)], called FLI, that we hypothesize improve preimplantation development of bovine embryos in vitro. To test this hypothesis, FLI was supplemented into oocyte maturation or embryo culture medium. Read More

View Article and Full-Text PDF
February 2021

Changes in telomere length and senescence markers during human ovarian tissue cryopreservation.

Sci Rep 2021 Jan 26;11(1):2238. Epub 2021 Jan 26.

Department of Obstetrics and Gynecology, Korea University College of Medicine, 73, Inchon-ro, Seongbuk-gu, Seoul, 02841, South Korea.

Ovarian tissue cryopreservation is considered as a useful option to preserve fertility for cancer patients. This study purposed to evaluate the change of telomere length and senescence markers during human ovarian tissue cryopreservation. Ovarian tissues were obtained from women who underwent benign ovarian surgery in the gynecology research unit of a university hospital with prior consent and IRB approval. Read More

View Article and Full-Text PDF
January 2021

Passive slow freezing is an efficacious and cost-effective alternative to controlled slow freezing for ovarian tissue cryopreservation.

Cryobiology 2021 Jun 21;100:164-172. Epub 2021 Jan 21.

Department for Reproductive Medicine, Ghent University Hospital, Ghent, Belgium. Electronic address:

We aimed to assess the feasibility of passive slow freezing (PSF using Mr. Frosty container, Nalgene) as an alternative to controlled slow rate freezing (CSF using (Freezal™, Air liquide)) for human ovarian tissue (OT) cryopreservation. Validation studies needed were determined after assessing the risk associated (EuroGTP-II ART tool) and were conducted in 66 OT samples from 10 transgender men aged 23. Read More

View Article and Full-Text PDF

Differential proteome between ejaculate and epididymal sperm represents a key factor for sperm freezability in wild small ruminants.

Cryobiology 2021 04 21;99:64-77. Epub 2021 Jan 21.

Department of Animal Reproduction, Spanish National Institute for Agricultural and Food Research and Technology (INIA), Avda Puerta de Hierro km 5.9, 28040, Madrid, Spain. Electronic address:

Epididymal sperm shows higher cryoresistance than ejaculated sperm. Although the sperm proteome seems to affect cell cryoresistance, studies aiming at identifying proteins involved in sperm freezing-tolerance are scarce. The aims of this study were to investigate differences of sperm freezability and proteome between epididymal and ejaculated sperm in three mountain ungulates: Iberian ibex, Mouflon and Chamois. Read More

View Article and Full-Text PDF