Freeze-induced shrinkage of individual cells and cell-to-cell propagation of intracellular ice in cell chains from salivary glands

Berger, W.; Uhrík, B

doi:10.1007/bf01938868

Cited by 59 publications

(42 citation statements)

References 22 publications

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“…The temperature at which 100% IIF occurred was higher in DPSC monolayers (−6°C) compared to DPSC suspensions (−12°C), which was consistent with previous observations that cell-to-cell contact increases the incidence of IIF [4–7,9,11,12]. Once IIF was achieved, cell preparation (suspension or monolayer) and final subzero temperature, to which cells were cooled, were both found to impact the incidence of membrane damage.…”

Section: Discussionsupporting

confidence: 88%

Intracellular ice formation in confluent monolayers of human dental stem cells and membrane damage

2010

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Dental pulp stem cells (DPSCs) are of interest to researchers and clinicians due to their ability to differentiate into various tissue types and potential uses in cell-mediated therapies and tissue engineering. Currently DPSCs are cryopreserved in suspension using Me(2)SO. However, preservation as two and three dimensional constructs, along with the elimination of toxic Me(2)SO, may be required. It was shown that intracellular ice formation (IIF), lethal to cells in suspensions, may be innocuous in cell monolayers due to ice propagation between cells through gap junctions that results in improved post-thaw recovery. We hypothesized that innocuous IIF protects confluent DPSC monolayers against injury during cryopreservation. The objective was to examine the effects of IIF on post-thaw viability of both confluent monolayers and suspensions of DPSCs. Confluent DPSC monolayers were assessed for the expression of gap junction protein Connexin-43. IIF was induced on the cryostage and in the methanol bath at different subzero temperatures. Membrane integrity and colony-forming ability were assessed post-thaw. Confluent DPSC monolayers expressed Connexin-43. In cell suspensions, 85.9+/-1.7% of cells were damaged after 100% IIF. In cell monolayers, after 100% IIF, only 25.5+/-5.5% and 14.8+/-3.3% of cells were damaged on the cryostage and in the methanol bath respectively. However, DPSC monolayers exposed to 100% IIF showed no colony-forming ability. We conclude that confluent monolayers of DPSCs express the gap junction-forming protein Connexin-43 and upon IIF retain membrane integrity, however lose the ability to proliferate.

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Section: Discussionsupporting

confidence: 88%

Intracellular ice formation in confluent monolayers of human dental stem cells and membrane damage

2010

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“…In a cluster of cells such an event may have more profound consequences. Cell-to-cell propagation of intracellular ice has been demonstrated in cultured cell monolayers [158] and cell strands from insect salivary glands [159]. It has been postulated that this occurs via gap junctions, and there is experimental evidence to support this.…”

Section: Mechanisms Of Slow Cooling Injurymentioning

confidence: 85%

“…The observation that intracellular ice propagates more readily between cells with gap junctions under certain conditions may at least in part explain the poor recovery of hES cells after slow cooling. Damage to the hESC clusters caused by intercellular ice propagation, either by random nucleation events within the cluster and propagation through the gap junctions [166] or from surface-catalysed nucleation at its periphery [167] followed by cell-to-cell propagation, could lead to disruption of the cell cluster affecting cell proliferation, differentiation and apoptosis on thawing [159]. Methods that attempt to control the nucleation event, such as seeding, may therefore act to initiate ice formation outside of the cellular (or cell cluster) compartment.…”

Section: Mechanisms Of Slow Cooling Injurymentioning

confidence: 99%

Cryopreservation of Human Stem Cells for Clinical Application: A Review

Hunt¹

2011

Transfus Med Hemother

275

207

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Stem cells have been used in a clinical setting for many years. Haematopoietic stem cells have been used for the treatment of both haematological and non-haematological disease; while more recently mesenchymal stem cells derived from bone marrow have been the subject of both laboratory and early clinical studies. Whilst these cells show both multipotency and expansion potential, they nonetheless do not form stable cell lines in culture which is likely to limit the breadth of their application in the field of regenerative medicine. Human embryonic stem cells are pluripotent cells, capable of forming stable cell lines which retain the capacity to differentiate into cells from all three germ layers. This makes them of special significance in both regenerative medicine and toxicology. Induced pluripotent stem (iPS) cells may also provide a similar breadth of utility without some of the confounding ethical issues surrounding embryonic stem cells. An essential pre-requisite to the commercial and clinical application of stem cells are suitable cryopreservation protocols for long-term storage. Whilst effective methods for cryopreservation and storage have been developed for haematopoietic and mesenchymal stem cells, embryonic cells and iPS cells have proved more refractory. This paper reviews the current state of cryopreservation as it pertains to stem cells and in particular the embryonic and iPS cell.

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“…Bartels and Potter (1993) have described communicating gap junctions between adjacent pavement cells of lamprey gills, and Sandbacka et al (1998) presented evidence for the movement of Lucifer yellow across gap junctions between adjacent rainbow trout (Oncorhynchus mykiss) gill epithelial cells grown in culture. Further studies examining the formation of intracellular ice in chains of insect salivary glands suggested that the mechanism of ice formation and the extent of cell damage could be influenced by gap junctions (Berger and Uhrik 1996). Such channels may render the cell membranes between adjacent pavement cells more permeable to ice propagation than the apical cell membranes.…”

Section: Discussionmentioning

confidence: 97%

Localization of cells from the winter flounder gill expressing a skin type antifreeze protein gene

Murray¹,

Hew²,

Kao³

et al. 2002

Can. J. Zool.

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In situ hybridization on whole mounts and paraffin-sectioned winter flounder (Pleuronectes americanus) gill, using riboprobes specific to a skin type antifreeze protein (AFP) gene, showed a mRNA distribution associated with cells throughout the filament and the lamellae. Immunohistochemistry using antibodies for a skin-type AFP identified cells corresponding to those detected using in situ hybridization. Parallel experiments with antibodies for chloride-cell markers showed that these cells were not involved in antifreeze-protein expression. Similarly, goblet cells did not show cross-reactivity with the AFP antibodies. This general distribution suggested that pavement cells were likely involved. Reverse transcription polymerase chain reaction using gill cDNA templates and subsequent sequencing of the products confirmed the presence of skin type AFP transcripts in this tissue. Expression of a AFP in this area may act as a first line of defence against ice-crystal migration into peripheral tissues.Résumé : L'hybridation in situ de montages entiers et de coupes à la paraffine de branchies de la plie rouge (Pleuronectes americanus) au moyen de ribo-sondes spécifiques au gène de la protéine antigel (AFP) de type cutané a démontré que la répartition de l'ARNm est associée aux cellules dans tout le filament et dans les lamelles. L'immuno-histo-chimie à l'aide d'anticorps de la protéine antigel (AFP) de type cutané a reconnu des cellules correspondant à celles obtenues par hybridation in situ. Des expériences en parallèle à l'aide d'anticorps de marqueurs des cellules à chlorures ont montré que ces cellules n'ont aucun rôle à jouer dans l'expression des protéines antigel. De même, les cellules caliciformes n'ont pas de réaction croisée avec les anticorps des protéines antigel. Cette répartition générale semble indiquer que les cellules pavimenteuses ont probablement un rôle à jouer. La transcription inverse-amplification en chaîne par polymérase au moyen de matrices d'ADNc des branchies et le séquençage subséquent des produits obtenus ont confirmé la présence de transcriptions d'AFP de type cutané dans ce tissu. L'expression des protéines antigel dans cette région peut servir de barrière à la migration de cristaux de glace dans les tissus périphériques.[Traduit par la Rédaction] 119 Murray et al.

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Freeze-induced shrinkage of individual cells and cell-to-cell propagation of intracellular ice in cell chains from salivary glands

Cited by 59 publications

References 22 publications

Intracellular ice formation in confluent monolayers of human dental stem cells and membrane damage

Intracellular ice formation in confluent monolayers of human dental stem cells and membrane damage

Cryopreservation of Human Stem Cells for Clinical Application: A Review

Localization of cells from the winter flounder gill expressing a skin type antifreeze protein gene

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