Cryopreservation of amniotic fluid-derived stem cells using natural cryoprotectants and low concentrations of dimethylsulfoxide.


:Amniotic fluid-derived stem cells (AFSCs) are a potential cell source for therapeutic applications. They can be easily mass produced, cryopreserved and shipped to clinics for immediate use. However, one major obstacle to the manufacturing of clinical grade stem cells is the need for current good manufacturing practices for cryopreservation, storage, and distribution of these cells. Most current cryopreservation methods used for stem cells include the potentially toxic cryoprotectant (CPA) dimethylsulfoxide (Me(2)SO) in the presence of animal serum proteins that prevent direct use of these cells in human therapeutic applications. To avoid any potential cryoprotectant related complications, it will be essential to develop non-toxic CPAs or reduce CPA concentration in the freezing media used. In this study, we assessed the use of disaccharides, antioxidants and caspase inhibitors for cryopreservation of AFSCs in combination with a reduced concentration of Me(2)SO. The thawed cells were tested for viability with MTT assays and a growth curve was created to measure population doubling time. In addition, we performed flow cytometry analysis for cell surface antigens, RT-PCR for mRNA expression of stem cell markers, and assays to determine the myogenic differentiation potential of the cells. A statistically significant (p<0.05) increase in post-thawed cell viability in solutions containing trehalose, catalase and (Z)VAD-fmk with 5% Me(2)SO was observed. The solutions containing trehalose and catalase with 5% or 2.5% (v/v) Me(2)SO produced results similar to those for the control (10% (v/v) Me(2)SO and 30% FBS) in terms of culture growth, expression of cell surface antigens and mRNA expression of stem cell markers in AFSCs cryopreserved for a minimum of 3 weeks. Thus, AFSCs can be cryopreserved with 1/4 the standard Me(2)SO concentration with the addition of disaccharides, antioxidants and caspase inhibitors. The use of Me(2)SO at low concentrations in cell freezing solutions may support the development of clinical trials of AFSCs.






Seo JM,Sohn MY,Suh JS,Atala A,Yoo JJ,Shon YH




Has Abstract


2011-06-01 00:00:00














  • Piglets born after vitrification of embryos using the open pulled straw method.

    abstract::Morulae and unhatched blastocysts from Large White hyperprolific (LWh) and Meishan (MS) gilts were selected to test an ultrarapid open pulled straw (OPS) vitrification method with two media. The viability of vitrified/warmed embryos was estimated by the percentage of embryos that developed to the hatched blastocyst st...


    pub_type: 杂志文章


    authors: Berthelot F,Martinat-Botté F,Locatelli A,Perreau C,Terqui M

    更新日期:2000-09-01 00:00:00

  • Viral contamination of embryos cryopreserved in liquid nitrogen.

    abstract::Despite the worldwide application of embryo-freezing technology as the means of preserving germplasm of mammalian species, there is no information available on the possible transmission of infectious agents to cryopreserved embryos via contaminated liquid nitrogen (LN). Recently, it has been reported that new methods ...


    pub_type: 杂志文章


    authors: Bielanski A,Nadin-Davis S,Sapp T,Lutze-Wallace C

    更新日期:2000-03-01 00:00:00

  • First steps towards Echinometra lucunter embryo cryopreservation.

    abstract::We have studied the sensitivity to cryoprotecting agents of different embryos of the local sea urchin, Echinometra lucunter which is the species used for embryo-larval bioassays in Brazil. We have located significant differences between both species sensitivity to cryoprotecting agents; while for P. lividus propylene ...


    pub_type: 杂志文章


    authors: Ribeiro MB,Furley T,Spago FR,Paredes E

    更新日期:2018-02-01 00:00:00

  • Restoration of resistance to osmotic swelling of vitrified mouse embryos by short-term culture.

    abstract::In cryopreservation of mammalian embryos, embryos can be injured by osmotic swelling during removal of the cryoprotectant after warming. We have shown that vitrified embryos are more sensitive to osmotic swelling than fresh cells but that sensitivity is reduced or abolished if vitrified cells are cultured for a short ...


    pub_type: 杂志文章


    authors: Edashige K,Asano A,An TZ,Kasai M

    更新日期:1999-06-01 00:00:00

  • Introduction to the special issue: Thermodynamic aspects of cryobiology.

    abstract::This brief paper introduces the subject of thermodynamics and the papers of the special issue on thermodynamic aspects of cryobiology. Thermodynamic terminology is defined for the non-specialist. ...




    authors: Elliott JA

    更新日期:2010-02-01 00:00:00

  • Strategies for exploration of freeze responsive gene expression: advances in vertebrate freeze tolerance.

    abstract::Winter survival for many cold-blooded species involves freeze tolerance, the capacity to endure the freezing of a high percentage of total body water as extracellular ice. The wood frog (Rana sylvatica) is the primary model animal used for studies of vertebrate freeze tolerance and current studies in my lab are focuse...


    pub_type: 杂志文章


    authors: Storey KB

    更新日期:2004-04-01 00:00:00

  • Cold-induced ultrastructural changes in bull and boar sperm plasma membranes.

    abstract::The effect of low temperatures on the ultrastructure of the plasma membrane of bull and boar spermatozoa was investigated. Cold-induced changes in the organization of sperm plasma membrane components were demonstrated by the use of fast-freezing combined with freeze-fracture electron microscopy. This preparation techn...


    pub_type: 杂志文章


    authors: De Leeuw FE,Chen HC,Colenbrander B,Verkleij AJ

    更新日期:1990-04-01 00:00:00

  • Hatching, survival and deformities of piracanjuba (Brycon orbignyanus) embryos subjected to different cooling protocols.

    abstract::Groups of one hundred Brycon orbignyanus embryos at the stage of blastopore closure were subjected to different cooling protocols. Different combinations and concentrations of cryoprotectants were tested: sucrose, methanol, ethylene glycol and dimethyl sulfoxide (Me2SO); at different temperatures (0.0±2.0°C and 8.0±2....


    pub_type: 杂志文章


    authors: Paes Mdo C,da Silva RC,do Nascimento NF,Valentin FN,Senhorini JA,Nakaghi LS

    更新日期:2014-12-01 00:00:00

  • Osmotic injury of PC-3 cells by hypertonic NaCl solutions at temperatures above 0 degrees C.

    abstract::Cell injury due to osmotic dehydration, which is regarded as a major cause of injury during freeze-thaw processes, was examined closely using a perfusion microscope. Human prostatic adenocarcinoma cells (PC-3), which were put in a chamber, were subjected to hyperosmotic stresses by perfusing NaCl solutions of varying ...


    pub_type: 杂志文章


    authors: Zawlodzka S,Takamatsu H

    更新日期:2005-02-01 00:00:00

  • Characterization and modulation of human mesenchymal stem cell stress pathway response following hypothermic storage.

    abstract::Human mesenchymal stem cell (hMSC) research has grown exponentially in the last decade. The ability to process and preserve these cells is vital to their use in stem cell therapy. As such, understanding the complex, molecular-based stress responses associated with biopreservation is necessary to improve outcomes and m...


    pub_type: 杂志文章


    authors: Corwin WL,Baust JM,Baust JG,Van Buskirk RG

    更新日期:2014-04-01 00:00:00

  • Environmental stress responsive expression of the gene li16 in Rana sylvatica, the freeze tolerant wood frog.

    abstract::Wood frogs (Rana sylvatica) can endure weeks of subzero temperature exposure during the winter with up to 65% of their body water frozen as extracellular ice. Associated with freezing survival is elevated expression of a number of genes/proteins including the unidentified gene, li16, first described in liver. The curr...


    pub_type: 杂志文章


    authors: Sullivan KJ,Storey KB

    更新日期:2012-06-01 00:00:00

  • Technique for cryopreservation of intestinal smooth muscle cells.

    abstract::Attempts were made to develop a simplified procedure for long-term cryopreservation of intestinal smooth muscle cells (ISMC). ISMC were collected from the ileum of Sprague-Dawley neonatal rats through cellular dissociation in trypsin. Cryopreservation method comprised of a rapid 1-step (protocol 1) and a slow 3-step (...


    pub_type: 杂志文章


    authors: Batista Lobo S,Denyer M,Gopalan RC,Javid FA

    更新日期:2008-10-01 00:00:00

  • Effects of cryopreservation on the immunogenicity of porcine arterial allografts in early stages of transplant vasculopathy.

    abstract:BACKGROUND:The number of revascularization procedures including coronary and lower extremity bypass, have increased greatly in the last decade. It suggests a growing need for vascular grafts. Cryopreserved allografts could represent a viable alternative but their immunologic reactivity remains controversial. METHODS:7...


    pub_type: 杂志文章


    authors: Solanes N,Rigol M,Khabiri E,Castellà M,Ramírez J,Roqué M,Agustí E,Roig E,Pérez-Villa F,Segalés J,Pomar JL,Engel P,Massaguer A,Martorell J,Rodríguez JA,Sanz G,Heras M

    更新日期:2005-10-01 00:00:00

  • Hypothermia protects H9c2 cardiomyocytes from H2O2 induced apoptosis.

    abstract::The purpose of our study was to investigate underlying basic mechanisms of hypothermia-induced cardioprotection during oxidative stress in a cardiomyocyte cell culture model. For hypothermic treatment we cooled H9c2 cardiomyocytes to 20°C, maintained 20min at 20°C during which short-term oxidative damage was inflicted...


    pub_type: 杂志文章


    authors: Diestel A,Drescher C,Miera O,Berger F,Schmitt KR

    更新日期:2011-02-01 00:00:00

  • Clinical utility of freeze-all approach in ART treatment: A mini-review.

    abstract::A significant proportion of couples at reproductive age rely on assisted reproductive technology to overcome infertility. In vitro fertilisation (IVF) involves typically the use of exogenous gonadotropins to stimulate the ovary to produce oocytes, which are collected surgically. After fertilization by conventional IVF...


    pub_type: 杂志文章,评审


    authors: Sciorio R,Esteves SC

    更新日期:2020-02-01 00:00:00

  • A 9 kDa antifreeze protein from the Antarctic springtail, Gomphiocephalus hodgsoni.

    abstract::A 9 kDA antifreeze protein (AFP) was isolated and purified from the Antarctic springtail, Gomphiocephalus hodgsoni. By combining selective sampling procedures and a modified ice affinity purification protocol it was possible to directly isolate a single AFP protein without recourse to chromatographic separation techni...


    pub_type: 杂志文章


    authors: Hawes TC,Marshall CJ,Wharton DA

    更新日期:2014-08-01 00:00:00

  • New approaches for studying the permeability of fish embryos: toward successful cryopreservation.

    abstract::This paper describes some new approaches for understanding the permeability of teleost embryos. The dechorionated zebrafish (Brachydanio rerio) was used as a model for basic studies of water and cryoprotectant permeability. These embryos are composed of two compartments, a large yolk (surrounded by the yolk syncytial ...


    pub_type: 杂志文章,评审


    authors: Hagedorn M,Hsu E,Kleinhans FW,Wildt DE

    更新日期:1997-06-01 00:00:00

  • Effect of cooling rates on the cold hardiness and cryoprotectant profiles of locust eggs.

    abstract::To examine the relationship between cooling rate and cold hardiness in eggs of the migratory locust, Locusta migratoria, the survival rates and cryoprotectant levels of three embryonic developmental stages were measured at different cooling rates (from 0.05 to 0.8 degrees C min(-1)) in acclimated and non-acclimated eg...


    pub_type: 杂志文章


    authors: Wang HS,Kang L

    更新日期:2005-10-01 00:00:00

  • Freezing monolayers of cells without gap junctions.

    abstract::Cells in monolayers have been reported to be more susceptible to freezing injury than the same cell type frozen in dispersed suspensions. There appears to be an enhanced susceptibility to intracellular freezing in the monolayers, which is thought to be facilitated by the presence of gap junctions allowing the spread o...


    pub_type: 杂志文章


    authors: Armitage WJ,Juss BK

    更新日期:2003-04-01 00:00:00

  • The effect of cryopreservation on ciliary beat frequency of human respiratory epithelium.

    abstract::The effect of cryopreservation on ciliary activity of human nasal respiratory epithelium was evaluated. Samples were cryopreserved in a solution containing nutrient medium, 10% fetal calf serum, and two different concentrations (10 or 20%) of dimethyl sulfoxide and stored in liquid nitrogen at -196 degrees C for 2 wee...


    pub_type: 杂志文章


    authors: Di Benedetto G,Gill J,Lopez-Vidriero MT,Clarke SW

    更新日期:1989-08-01 00:00:00

  • Effect of the desiccation tolerance and cryopreservation methods on the viability of Citrus limon L. Burm cv. Eureka seeds.

    abstract::Cryopreservation of the germplasm for long-term periods is of great importance to maintain the genetic resource. Argentina is one of the world's highest lemon producing country. The performance of different cooling/warming rates in the cryopreservation method of Citrus limon L. Burm cv. Eureka seeds and their influenc...


    pub_type: 杂志文章


    authors: Orjuela-Palacio JM,Graiver N,Santos MV,Zaritzky NE

    更新日期:2019-08-01 00:00:00

  • Non-transmission of bacterial and viral microbes to embryos and semen stored in the vapour phase of liquid nitrogen in dry shippers.

    abstract::Cryopreservation and storage of germplasm is an important factor in the prevention of disease transmission by embryo transfer and artificial insemination. Here we report the results of an investigation on transmission of selected bacterial and viral pathogens by the vapour phase of liquid nitrogen (VPLN) to embryos an...


    pub_type: 杂志文章


    authors: Bielanski A

    更新日期:2005-04-01 00:00:00

  • Gas exchange measurement as a non-destructive viability assay for frozen-thawed, winter-dormant apple buds.

    abstract::Low temperature studies with winter-dormant buds are severely limited by the lack of a rapid,non-destructive assay for their viability. Investigations involving the winter harvest of ecodormant buds of woody subjects, including cryopreservation, are restricted if viability cannot be assessed until dormancy is broken. ...


    pub_type: 杂志文章


    authors: Vogiatzi C,Rosenqvist E,Grout BWW

    更新日期:2018-05-06 00:00:00

  • Vitrification of in vitro produced goat blastocysts: effects of oocyte donor age and development stage.

    abstract::This study examines the effectiveness of the cryotop vitrification method for the cryopreservation of goat blastocysts. To determine the effects of embryo development stage and donor age on in vitro survival rates, good-quality blastocysts from adult and prepubertal goats were sorted into non-expanded, expanded, hatch...


    pub_type: 杂志文章


    authors: Morató R,Romaguera R,Izquierdo D,Paramio MT,Mogas T

    更新日期:2011-12-01 00:00:00

  • Vitrification of mature mouse oocytes in a 6 M Me2SO solution supplemented with antifreeze glycoproteins: the effect of temperature.

    abstract::Oocytes have been successfully cryopreserved using rapid and slow freezing procedures. However, variability in the success of replicates has limited its practical application. In the present study, mature mouse oocytes were vitrified in 6 M dimethyl sulfoxide supplemented with 1 mg/ml antifreeze glycoproteins (AFGP) (...


    pub_type: 杂志文章


    authors: O'Neil L,Paynter SJ,Fuller BJ,Shaw RW,DeVries AL

    更新日期:1998-08-01 00:00:00

  • Machine perfusion enhances hepatocyte isolation yields from ischemic livers.

    abstract:BACKGROUND:High-quality human hepatocytes form the basis of drug safety and efficacy tests, cell-based therapies, and bridge-to-transplantation devices. Presently the only supply of cells derives from an inadequate pool of suboptimal disqualified donor livers. Here we evaluated whether machine perfusion could ameliorat...


    pub_type: 杂志文章


    authors: Izamis ML,Perk S,Calhoun C,Uygun K,Yarmush ML,Berthiaume F

    更新日期:2015-10-01 00:00:00

  • Insights into the model of non-perfect osmometer cells for cryopreservation: A parametric sweep analysis.

    abstract::Recently, a mathematical model able to describe the non-perfect osmotic behavior of cells during cryopreservation was proposed. The model improves the two-parameter formalism typically adopted in cryopreservation literature by allowing the transmembrane permeation of ions/salt, through the temporary opening of mechano...


    pub_type: 杂志文章


    authors: Traversari G,Cincotti A

    更新日期:2020-11-27 00:00:00

  • The effects of rapid cooling (cold shock) of ram semen, photoperiod, and egg yolk in diluents on the survival of spermatozoa before and after freezing.

    abstract::The effects of rapid cooling of semen (cold shock) from 30 degrees C to various temperatures above 0 degrees C on survival of ram spermatozoa suspended in diluents with or without egg yolk were assessed before and after freezing. Rapid cooling of extended semen from 30 to 15 degrees C had little or no effect on sperma...


    pub_type: 杂志文章


    authors: Fiser PS,Fairfull RW

    更新日期:1986-12-01 00:00:00

  • Influence of cell concentration on the contribution of unfrozen fraction and salt concentration to the survival of slowly frozen human erythrocytes.

    abstract::We have shown previously that the survival of human red blood cells during slow freezing at 2% hematocrit is dependent more on the magnitude of the unfrozen fraction than on the salt concentration in that unfrozen fraction. In parallel, first Nei and more recently Pegg and colleagues have shown that survival is affect...


    pub_type: 杂志文章


    authors: Mazur P,Cole KW

    更新日期:1985-12-01 00:00:00

  • Cryohemostasis of uncontrolled hemorrhage from liver injury.

    abstract::Uncontrolled hemorrhage is the primary cause of death in both blunt and penetrating liver trauma. Cryohemostasis was attempted in the past for elective liver surgery but did not gain popularity. During past decades, cryoequipment was refined and successfully used for tumor ablation. The purpose of the present study wa...


    pub_type: 杂志文章


    authors: Kopelman D,Klein Y,Zaretsky A,Ben-Izhak O,Michaelson M,Hashmonai M

    更新日期:2000-05-01 00:00:00