CryoLetters Logo
CryoLetters Logo
Abstracts: CryoLetters 31 (1), 2010

 

 

Volume 31, No. 1 January/February 2010

ISSN 0143-2044

 

 


Detection of 8-hydroxy-2'-deoxyguanosine as a marker of oxidative damage in dna and germplasm exposed to cryogenic treatments
Jason W. Johnston, Isobel Pimbley, Keith Harding and Erica E. Benson

1-13

 

 


Cryopreservation of Thymus moroderi by droplet vitrification
Ana Marco-Medina, José Luis Casas, Rony Swennen and Bart Panis

14-23

 

 


Vitrification of mosses: a useful method for the cryopreservation of Splachnum ampullaceum Hedw.
Rubén Mallón, Juan Rodríguez-Oubiña and María Luz González

24-28

 

 


Critical moisture content windows differ for the
cryopreservation of pomelo (Citrus grandis) seeds and embryonic axes

Bin Wen, Chuantao Cai, Ruling Wang, Yunhong Tan and Qinying Lan

29-39

 

 


Cryopreservation of fraser photinia (Photinia x fraseri Dress.) via vitrificationbased one-step freezing techniques
Yelda Ozden Tokatli and Hülya Akdemir

40-49

 

 


Effect of timing, dose and interstitial versus nanoparticle delivery of tumor necrosis factor alpha in combinatorial adjuvant cryosurgery treatment of elt-3 uterine fibroid tumor
Jing Jiang and John Bischof

50-62

 

 


Cryopreservation of Fraxinus excelsior L. embryogenic callus by one-step freezing and slow cooling techniques
E. A.Ozudogru, M. Capuana, E. Kaya, B. Panis and M. Lambardi

63-75

 

 


Abstracts to a Meeting of Workgroup 2 of the COST Action 871 ("Cryopreservation of Crop Species in Europe"). The title of the meeting was "Integration of Cryopreservation in Genebank Strategies". It was held at IPK Gatersleben, Germany, on September 11 09 2009.

76-94

 

 

 

Top of page

CryoLetters 31 (1), 1-13 (2010)
© CryoLetters,
businessoffice@cryoletters.org

DETECTION OF 8-HYDROXY-2'-DEOXYGUANOSINE AS A MARKER OF OXIDATIVE DAMAGE IN DNA AND GERMPLASM EXPOSED TO CRYOGENIC TREATMENTS

Jason W. Johnston1,2*, Isobel Pimbley1, Keith Harding3 and Erica E. Benson3

1University of Abertay Dundee, Bell Street, Dundee DD1 1HG, Scotland, UK.
2Current address: The New Zealand Institute for Plant and Food Research Ltd, 120 Mt Albert Rd, Private Bag 92 169, Mt Albert, Auckland, New Zealand.
3Research Scientists, Damar, Drum Road, Cuparmuir, Fife, KY15 5RJ, Scotland, UK.
*Corresponding author  email:
JJohnston@hortresearch.co.nz

Abstract

An HPLC method has been optimised to measure 8-hydroxy-2'-deoxyguanosine (8OHdG) in DNA and germplasm with the objective of using the adduct as a marker of cryostorage stability. The encapsulation-dehydration cryopreservation protocol was adapted as a model system for assessing the formation of 8OHdG from alginate-encapsulated DNA of calf thymus (CT) and currant species (Ribes) exposed to temperatures of -20 and -196ºC. The presence of H2O2 exacerbated the formation of 8OHdG in encapsulated CT and Ribes DNA. Production of the oxidized adduct was lower in the plant system. A reduction in residual water following osmotic dehydration and evaporative desiccation was associated with reduced adduct formation in encapsulated DNA. No significant differences in 8OHdG adduct formation were observed in plants regenerated from cryopreserved Ribes meristems derived from genotypes known to have differential tolerance to cryopreservation.

Keywords: cryobiology, DNA, free radicals, oxidative stress, 8OHdG, stability

 

 

Top of page

CryoLetters 31 (1), 14-23 (2010)
© CryoLetters,
businessoffice@cryoletters.org

CRYOPRESERVATION OF Thymus moroderi BY DROPLET VITRIFICATION

Ana Marco-Medina1, José Luis Casas1*, Rony Swennen2 and Bart Panis2

1Laboratory of Plant Biotechnology, Institute of Biodiversity CIBIO (Centro Iberoamericano de la Biodiversidad), University of Alicant, Crta. San Vicente del Raspeig s/n. E-03690 San Vicente del Raspeig, Alicante (Spain).
2Laboratory of Tropical Crop Improvement, Division of Plant Biotechnics, K.U. Leuven, Kasteelpark Arenberg 13, B-3001 Leuven, Belgium
*Corresponding author  e-mail:
jl.casas@ua.es

Abstract

Thymus moroderi Pau ex Martínez (Labiatae) was successfully cryopreserved using the droplet vitrification method. After 20 min in loading solution at room temperature, shoot tips were dehydrated with PVS2 at 0ºC for 30 min and immersed into LN. For thawing, shoot-tips were transferred into recovery solution for 15 min. A test of different recovery media revealed that the best results were obtained when the medium was supplement with 0.275 µM BA.

Keywords: cryopreservation, droplet vitrification, hyperhydration, in vitro conservation, Thymus moroderi

 

 

Top of page

CryoLetters 31 (1), 24-28 (2010)
© CryoLetters,
businessoffice@cryoletters.org

VITRIFICATION OF MOSSES: A USEFUL METHOD FOR THE CRYOPRESERVATION OF Splachnum ampullaceum Hedw.

Rubén Mallón1,3, Juan Rodríguez-Oubiña2 and María Luz González1*

1 Departamento de Fisiología Vegetal, Universidad de Santiago de Compostela, 15782 Santiago de Compostela, Spain.
2 Departamento de Botánica, Universidad de Santiago de Compostela, 15782 Santiago de Compostela, Spain.
3
Current address: Instituto de Investigaciones Agrobiológicas de Galicia (CSIC), 15780 Santiago de Compostela, Spain.
*Corresponding author  e-mail:
mluz.gonzalez@usc.es

Abstract

The source of germplasm as well as the technique used for storage of mosses can enhance survival after cryopreservation. Samples of gametophores, protonemata and protonemal brood cells from in vitro cultures of Splachnum ampullaceum were cryopreserved following exposure to a plant vitrification solution (PVS2) for two different times (5 and 10 min) at 0ºC. Half of the samples were pretreated with a loading solution containing 2 M glycerol and 0.4 M sucrose before exposure to PVS2. After one week storage in liquid nitrogen, S. ampullaceum samples were regenerated on Gamborg's B5 mineral medium with B5 vitamins. Exposure to a loading solution was a prerequisite for high survival in all samples. Four weeks after cryopreservation, 92.3% brood cells, 60.0% gametophores and 46.0% protonemata pretreated with a loading solution had regenerated, displaying normal growth and development, thus demonstrating that vitrification is a useful method for moss cryopreservation.

Keywords: bryophyte, moss, cryopreservation, vitrification, brood cells.

 

 

Top of page

CryoLetters 31 (1), 29-39 (2010)
© CryoLetters,
businessoffice@cryoletters.org

CRITICAL MOISTURE CONTENT WINDOWS DIFFER FOR THE CRYOPRESERVATION OF POMELO (Citrus grandis) SEEDS AND EMBRYONIC AXES

Bin Wen*, Chuantao Cai, Ruling Wang, Yunhong Tan and Qinying Lan

Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Mengla, Yunnan, 666303, China.
*Correspondence author  e-mail:
wenb@xtbg.org.cn

 Abstract

Cryopreservation was attempted using a partial dehydration and freezing protocol on pomelo (Citrus grandis) seeds of ten cultivars and embryonic axes of one cultivar collected from Xishuangbanna. Although seeds of all ten cultivars could be dehydrated safely to 10% moisture content, further dehydration impaired seed viability with critical moisture contents ranging from 9% to 7%. Complete seedling regenerated from seeds frozen in a moisture window between 5-9%, and maximum seedling recovery varied between 22-86%. Cryopreservation of Jiajieyou cv. embryonic axes had a moisture window between 3-20%, much wider compared to whole seed cryopreservation, and maximum post-thaw emergence of 93% was achieved at 13% moisture content. Emergence differed only slightly from survival when whole seeds were cryopreserved, but for the embryonic axis cryopreservation at low moisture contents resulted in much lower emergence than survival. Possible causes of intraspecific variation in cryotolerance in pomelo seeds is discussed.

Keywords: Citrus grandis; cryotolerance; desiccation tolerance; intermediate seeds; intraspecific variation

 

 

Top of page

CryoLetters 31 (1), 40-49 (2010)
© CryoLetters,
businessoffice@cryoletters.org

CRYOPRESERVATION OF FRASER PHOTINIA (Photinia x fraseri Dress.) VIA VITRIFICATION-BASED ONE-STEP FREEZING TECHNIQUES

Yelda Ozden Tokatli* and Hülya Akdemir

Gebze Institute of Technology, Faculty of Science, Department of Biology, 41400, Kocaeli, Turkey
*Corresponding author  e-mail:
yelda75@yahoo.com, ozden@gyte.edu.tr

Abstract

An efficient vitrification-based cryopreservation procedure was developed for Fraser photinia shoot apices by assessing the influences of various vitrification solutions (PVS1, PVS2, PVS3 and VSL) and different vitrification methods (cryovial vitrification, droplet vitrification and droplet freezing) on shoot regrowth. Moreover, influences of cold-hardening period (0 to 8 weeks), preculture medium (with sucrose and proline) and regrowth medium (QL plus 4.4, 8.8 and 17.6 µM BA) were also evaluated. Among the different procedures tested, best shoot regrowth (40.3%) was achieved by using a droplet vitrification technique in which cold-hardened and precultured shoot apices were vitrified for 120 min at 0°C in droplets, rapidly cooled, thawed and then cultured on 17.6 µM BA-containing QL medium. Overall results indicated the importance of not only the composition of vitrification solution, and preculture and regrowth media, but also the application of an appropriate vitrification technique to achieve optimum recovery post-cryopreservation. 

Keywords: cryovial vitrification, droplet vitrification, PVS1, PVS2, PVS3, VSL

 

 

Top of page

CryoLetters 31 (1), 50-62 (2010)
© CryoLetters,
businessoffice@cryoletters.org

EFFECT OF TIMING, DOSE AND INTERSTITIAL VERSUS NANOPARTICLE DELIVERY OF TUMOR NECROSIS FACTOR ALPHA IN COMBINATORIAL ADJUVANT CRYOSURGERY TREATMENT OF ELT-3 UTERINE FIBROID TUMOR

Jing Jiang1 and John Bischof*1,2,3

Department of Biomedical Engineering1, Mechanical Engineering2 and Urologic Surgery3, University of Minnesota, Minneapolis, MN 55455, USA.
*Corresponding author email:
bischof@umn.edu

Abstract

Cryosurgery has shown potential as a minimally invasive technology for tumor treatment. However, incomplete destruction followed by tumor recurrence after cryosurgery is a common drawback. This study characterizes several variables in the cryoadjuvant TNF-α enhancement of conservative cryosurgery (i.e. freezing to the visible edge) of ELT-3 (uterine leiomyoma) tumor in a female nude mouse model. The variables include pretreatment time, mode of TNF-α delivery (native vs. CYT-6091, a PEGylated 33 nm colloidal gold core nanoparticle) and dose of TNF-α. Survival and tumor growth delay were measured up to 30 days and showed: 1) pretreatment with TNF-α required 4 hours incubation prior to cryosurgery to produce a tumor growth delay over cryosurgery alone, and 2) CYT-6091 reduced the toxicity of TNF-α administration over intratumoral or peritumoral injection of native TNF-α. Taken together, 5 µg TNF-α delivered by the nanodrug CYT-6091 4 hours prior to cryosurgery yielded a dramatic reduction in tumor growth over cryosurgery alone and in some cases even total remission of the tumor. However, some toxicity at higher doses (i.e. 5 µg) with CYT-6091 was noted compared to previous work in prostate (LNCaP) cancer grown in a male nude mouse. Potential reasons for this, including sex and weight of the animals are discussed. Further opportunities to optimize the TNF-α enhanced cryosurgical response of this tumor include dosing between 2 - 5 µg at 4 hours prior to cryosurgery, and freezing beyond the visible edge of the tumor.

Keywords: Cryosurgery, TNF-α, CYT-6091, leiomyoma, drug delivery

 

 

Top of page

CryoLetters 31 (1),63-75 (2010)
© CryoLetters,
businessoffice@cryoletters.org

CRYOPRESERVATION OF Fraxinus excelsior L. EMBRYOGENIC CALLUS BY ONE-STEP FREEZING AND SLOW COOLING TECHNIQUES

E. A.Ozudogru1, M. Capuana2, E. Kaya1, B. Panis3 and M. Lambardi4*

1GYTE / Gebze Yuksek Teknoloji Enstitusu, Istanbul Caddesi, No 101, 41400, Gebze, Kocaeli, Turkey
2IGV / Istituto di Genetica Vegetale, CNR, Via Madonna del Piano 10, 50019, Sesto Fiorentino, Florence, Italy
3Laboratory of Tropical Crop Improvement, K.U. Leuven, Kasteelpark Arenberg 13, 3001, Leuven, Belgium
4IVALSA / Istituto per la Valorizzazione del Legno e delle Specie Arboree, CNR, Via Madonna del Piano 10, 50019, Sesto Fiorentino, Florence, Italy
*Corresponding author  email:
lambardi@ivalsa.cnr.it

Abstract

An efficient cryopreservation protocol for the safe storage of Fraxinus excelsior L. embryogenic callus cultures is reported. The cryopreservation methods tested included one-step freezing by means of (i) encapsulation-vitrification; or (ii) encapsulation-dehydration; and (iii) slow cooling using the Nalgene Freezing container, Mr Frosty®, which produces a temperature decrease of about 1ºC min-1 when placed in a -70ºC freezer. None of the one-step freezing techniques was effective for cryopreservation of encapsulated callus masses, irrespective of the cryoprotective treatment applied, i.e., treatment with the PVS2 vitrification solution or physical dehydration with silica gel before direct immersion in liquid nitrogen. On the contrary, when a slow cooling protocol was applied to embryogenic callus which had been pretreated for 60 min with a 210 g l-1 (0.61 M) sucrose-7.5% DMSO cryoprotective solution, up to about 1.3 g per Petri dish of proliferating callus was observed 42 days after recovery from liquid nitrogen, and cultures were able to produce somatic embryos 8 weeks after transfer to semi-solid medium. TTC staining of callus cultures provided a fast evaluation of culture viability.

Keywords: common ash, encapsulation-dehydration, encapsulation-vitrification, slow cooling, somatic embryogenesis

CryoLetters Logo
CryoLetters Logo

Home  Aims and Scope  Abstracts  Editorial Board  Info for Authors  Subscriptions  Links

Please contact CryoLetters with questions or comments.
© Copyright 2000-2012 CryoLetters.  All rights reserved.

Site updated: 07 May, 2017

Abstracts

Volume 38 (2017)
Volume 37 (2016)
Volume 36 (2015)
Volume 35 (2014)
Volume 34 (2013)
Volume 33 (2012)
Volume 32 (2011)
Volume 31 (2010)
Volume 30 (2009)
Volume 29 (2008)
Volume 28 (2007)
Volume 27 (2006)
Volume 26 (2005)
Volume 25 (2004)
Volume 24 (2003)
Volume 23 (2002)
Volume 22 (2001)
Volume 21 (2000)
Volume 20 (1999)

For Abstracts published from meetings, such as SLTB meetings, go to the relevant Volume Year  of the journal (above).
Abstracts are often published by the journal in the Year subsequent to the Meeting's Date

For Full text Free Access Content (from 2000 onwards) go to CryoLetters at Ingenta and look for the blue symbol.