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1

Cryopreservation of plant tissue culture

100%
Makowska Z., Kotlinska T.
Biotechnologia
|
2001
|
issue 2
126-134
EN
Three methods of cryopreservation: vitrification, encapsulation and slow freezing were discussed. The factors having influence on long-term storage in liquid nitrogen as selection of plant explants, media, optimal condition for regeneration and growth of plants were described. Practical utilisation of vitrification method and obtained results on garlic (Allium sativum L.) are given as an example. Vitrification involves treatment of samples with cryoprotective substances, dehydration with highly concentrated vitrification solution, aimed at water removal from the cells in order to protect them against destruction because of liquid nitrogen influence. This method offers a possibility to store plant material for a long time without any modification and contamination. The aim of the study is to develop a useful method for cryopreservation of apical meristems of garlic (Allium sativum L.) bulbils. The apical meristems of garlic bulbils were treated with PVS 3 solution containing 50% sucrose and 50% glicerol. The time of treatment of explants with PVS 3 was from 0 min to 240 min, then samples were plunged into liquid nitrogen (LN2) where they were kept for 1 hour or 30 days. Rapid heat up was achieved by plunging the samples in a 45oC water-bath until cryoprotectant solution became liquid. After cryopreservation, survival and regrowth on MS (Murashige and Skoog) medium were observed. Results showed that the best kind of explants giving high percentage of regeneration were meristems with two leaves primordia about 1 mm of diameter and about 3 mm in length.
2

Cryopreservation of plant cell culture

100%
van Iren F., Schrijnemakers E., Reinhoud P.
Biotechnologia
|
1996
|
issue 1
60-68
EN
Some relatively simple and cheap procedures for long term preservation in liquid nitrogen have been worked out.These are based both on two-step freezing (in which the specimen is progressively dehydrated when ice crystals grow in the medium during slow cooling) and on vitrification (absence of ice crystals due to rapid freezing in highly concentrated vitrifying solutions).For recalcitrant strains alternative storage methods have been studied.
3

Cryopreservation efficacy in protection of (epi)genetic stability of plant material

100%
Mikula A.
Biotechnologia
|
2010
|
issue 2
23-37
EN
Cryopreservation is important for the long-term conservation of plant genetic resources, especially for the species producing recalcitrant seeds and clonally propagated crops which are proliferated through grafting or as vegetative cuttings, suckers, roots, tubers and bulbs. Despite the fact that in liquid nitrogen temperature the cell division and metabolism are arrested, the factors associated with cryotreatment, cryostorage or plant recovery could be a source of somaclonal variations. The lecture assesses an attempt of attained knowledge within the influence of cryopreservation on the genetic integrity of regenerated plants or recovered tissue. Assessment of genetic and epigenetic stability of the recovered plants derived from cryopreserved plant material is an important step to success of any storage protocol. Until now, from among 64 published papers, only 7 have shown genetic changes in plantlets regenerated after cryopreservation, and 3 - variability in tissue recovered after cryostorage without consequences in regenerants. Epigenetic changes were described using isoschisomers and MSAP, AMP or RAPD methods in 9 works. It was suggested that the processes of cryoprotection and cryostorage had an impact on DNA methylation status, it could lead to alterations in chromatin structure and changes in gene expression. However, majority of the works reported on the insignificant or any influence of cryopreservation on the plant material.
4

Cryopreservation of suspended cells of selected Gentiana

100%
Mikula A., Rybczynski J. J.
|
2003
|
issue 3
39-51
EN
Cryopreservation is recognised as the most efficient method of plant genome preservation and embryogenic potential of plant in vitro cultures with non-changed status. Because of the risk of biological degradation of tissue during freezing treatment and it various sensibility on the chilling stress, the individual treatment with cryoprotectance and parameters of the temperature decreasing is highly required. In this presentation, following subjects will be discussed: 1) description of suspension cultures of various gentians, 2) cell pretreatments for surviving the stress of low temperature, 3) rearrangement of cell cytoplasm because of high sucrose concentrations, 4) the role of sucrose in the development of freezing tolerance and surviving of gentaiana suspensions, 5) the effect of strong plasmolysis induced by 3 M glucose.
5

Characterization of human hepatocytes isolated from non-transplantable livers

100%
Laba A., Ostrowska A., Patrzalek D., Paradowski L., Lange A.
Archivum Immunologiae et Therapiae Experimentalis
|
2005
|
vol. 53
|
issue 5
442-453
EN
The successful use of hepatocytes depends on a reliable demonstration of the functional and morphological integrity of isolated cells. Herein we investigated whether the isolation and cryopreservation of primary human hepatocytes can compromise cell viability and liver-specific characteristics. Hepatocytes were isolated from encapsulated human liver segments by a modified 2-step perfusion technique. Isolated cells were Percoll-purified, cryopreserved, and stored in liquid nitrogen for 1?12 months. For rapid assessment of fresh and cryopreserve/thawed hepatocyte yield and viability, the cells were stained with trypan blue or labeled with fluorochromes. For immunocytochemical analysis, the cells were labeled with monoclonal antibodies for the presence of the following antigens and chemokines: CD3, CD45Ro, CD45Ra, CD34, CD68, CD90, CD95, CD20, HLA-DR, Ki67, PCNA, Bcl-2, p53, CXCR3, CXCR4, and SDF-1. The cells were tested for several specific functions, such as ureagenesis, energy status, MTT activity, lactate dehydrogenase leakage, and total CYP450 content. Assessment of both freshly isolated (Percoll-purified) and cryopreserved/thawed hepatocytes revealed a low constitutive level of contamination by non-parenchymal cells compared with crude (unpurified) preparations and tissue sections. All viable hepatocytes showed intact morphology and retained CYP450 protein, energy status, and urea synthesis. Modifications in hepatocyte preparations, such as depletion of dead, damaged, and non-parenchymal cells, improves cell purity, which can be adapted to further evaluation of hepatocyte immunogenicity. These data illustrate the importance and feasibility of human hepatocyte banking.
6

Cryopreservation - a tool for long-term storage of cells, tissues and organs from in vitro culture derived

88%
Mikula A., Rybczynski J. J.
Biotechnologia
|
2006
|
issue 4
145-163
EN
Cryopreservation offers the possibility for long-term storage of genetic resources with maximal genotypic stability, using a minimum of space and maintenance. At present it is actively used all over the world for storage of plant material: seeds, pollen, spores, dormant buds or apical meristems in genebanks. The development of biotechnology led to the production of a new category of germplasm for cryostorage: in vitro obtain tissues, organs, embryos, special cell lines and genetically modified plant material. The maintenance of in vitro collections remains risky regarding losing accessions due to the contamination, human error or somaclonal variation. The classical slow cooling was the first standard protocol developed for hydrated plant tissues. This method is mainly used for cryopreservation of non-organized tissues, for example: cell suspensions and calli, or apices of cold-tolerant species. For differentiated structures, new cryopreservation techniques such as vitrification and encapsulation/dehydration procedures or droplet method are efficient and reliable. These freezing techniques have been successfully, routinely applied for cryopresevation of various plant material of temperate and tropical climate species. So far, cryopreservation procedures are developed for in vitro tissues and recalcitrant seeds of about 100 and 40 species, respectively.
7

Methods of preservation of in vitro plant cell culture

88%
Marecik R., Kroliczak P.
Biotechnologia
|
1998
|
issue 1
105-116
EN
Tissue culture has been utilized for producing virus free plants for vegetative propagation and genetic engineering. In addition, altered cells lines showing higher productivity of secondary metabolities can be obtained using tissue culture. The wide use of tissue culture requires the development of new preservation techniques for in vitro culture material. One of this method is growth reduction achieved by modifying various parameters such as: temperature, culture medium, gasous environment. However, crypreservation (i. e. storage in liquid nitrogen. -196?C) is the only method available nowadays for long-term conservation. New cryopreservation techniques such as encapsulation-dehydratation, vitrification and desiccation helped expend the list of plant species that can tolerate low temperatures and are characterized by a normal rate of growth. Each step of the cryopreservation procces requires specific conditions.
8

Cryopreservation as a method to facilitate preservation of life capacity of plant cells and tissues

88%
Mikula A.
Biotechnologia
|
2008
|
issue 2
41-57
EN
This paper presents a review of fundamental aspects of plant cryopreservation. Liquid nitrogen has several advantages over storage of vegetatively propagated material under normal low-temperature in vitro culture and could also help in preserving genetic biodiversity. Development of efficient cryopreservation protocols based on the induction of tolerance to freezing and/or desiccation is also discussed. Cold and/or preculture acclimatization leads to ultrastructural, physiological and molecular changes in cells and they are important for improving viability after cryopreservation. The application of vitrification-based procedures and ultra-fast freezing/thawing rates could be effective and reliable for wide variety of plant species/ tissues and relatively genotype independent. Majority of papers demonstrate that the liquid nitrogen allows high viability rates and re-growth without a loss of biosynthetic capacity. Up to now, there has been no clear evidence of morphological, cytological or genetic alterations due to cryopreservation.
9

Induction, proliferation and maintenance of embryogenic tissues of Serbian spruce in liquid nitrogen

75%
Hazubska-Przybyl T., Bojarczuk K.
Biotechnologia
|
2010
|
issue 2
96-104
EN
Picea omorika (Pancic.) Purk. is an endemic species, threatened with extinction in the wild. Besides, it is a valuable species for nursery production. Hence it is necessary to protect its genetic resources ex situ. Somatic embryogenesis is a very beneficial micropropagation method because of potentially high regeneration rate and possibilities to maintain embryogenic tissue and somatic embryos in liquid nitrogen (LN). That is why the aim of this study was to induce embryogenic tissue culture of Picea omorika (Pancic.) Purk.
10

Cryopreservation of porcine oocytes and embryos

75%
Gajda B., Smorag Z.
|
2003
|
issue 1
116-228
EN
This paper presents the current possibilities, state of knowledge and prospects for cryopreservation of pig oocytes and embryos. The main factors of cryopreservation efficiency, methods for the evaluation of cryopreserved embryos, and the possibilities of modifying their susceptibility to cryopreservation are discussed. In addition, the most significant results of pig embryo freezing and vitrification and the cryotechnical aspects of this method are presented.
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