Cryopreservation of Anchorage-Dependent Mammalian Cells Fixed to Structured Glass and Silicon Substrates

Hornung, Jan; Müller, Torsten; Fuhr, G.

doi:10.1006/cryo.1996.0026

Cited by 29 publications

(17 citation statements)

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“…These facts suggest that heat and mass transfer within the porous scaffold were sufficient in this study. It has been reported that 60–75% of viable cells were recovered after thawing cryopreserved fibroblasts that were attached to a glass chip, a silicon chip, or a polyglycolic acid scaffold (9,18). Compared with these results, a similar or slightly higher recovery rate was obtained in the present study (Fig.…”

Section: Discussionsupporting

confidence: 82%

Cryopreservation of Fibroblasts Immobilized Within a Porous Scaffold: Effects of Preculture and Collagen Coating of Scaffold on Performance of Three‐Dimensional Cryopreservation

et al. 2010

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As a preliminary investigation to establish a cryopreservation method suited for bioartificial livers (BALs), three-dimensional (3-D) cryopreservation experiments with fibroblasts were performed, in which the cells were firstly seeded into a porous scaffold, and the scaffold containing the cells was then cryopreserved. After thawing, 65% of the initially applied cells were still attached to the scaffold, and this efficiency was significantly higher than that in the control experiments (39%), in which fibroblasts cryopreserved in a suspension were seeded into the scaffold. This higher efficiency was mainly caused by higher immobilization efficiency at the time of cell seeding (83%) than in the controls (54%). Collagen coating of the scaffold in the 3-D cryopreservation enhanced immobilization efficiency at the time of cell seeding, and 1-day precultures before the 3-D cryopreservation considerably improved cell growth after thawing. From these favorable results, this 3-D cryopreservation method may become useful for developing BALs.

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Section: Discussionsupporting

confidence: 82%

Cryopreservation of Fibroblasts Immobilized Within a Porous Scaffold: Effects of Preculture and Collagen Coating of Scaffold on Performance of Three‐Dimensional Cryopreservation

et al. 2010

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“…Almost all previous investigations of the effect of cell-cell interactions on the IIF process have compared freshly trypsinized cells in suspension, or single cells adherent to glass, with cells of the same type cultured in confluent monolayers on glass (e.g., Porsche et al, 1991;Armitage and Juss, 1996;Acker and McGann, 2000), or suspension cultured in aggregates (e.g., McGann et al, 1972;McGrath et al, 1975;Acker et al, 1999). However, previous experimental designs have been unable to control confounding effects resulting from cell damage by enzymatic digestion (McGann et al, 1972;Sandler and Andersson, 1984;Yarmush et al, 1992), from changes in phenotype due to time in culture (Hetzel et al, 1973;Darr and Hubel, 2001) or cell attachment and spreading (Hornung et al, 1996;Chen et al, 1997), or from mass transport limitations in cell clusters (Levin et al, 1977). Whereas all aforementioned factors affect IIF and cryoinjury, past attempts to attribute differences in probability of IIF or cell survival for different culture configuration to hypothesized effects of cell-cell contact have not been conclusive.…”

Section: Discussionmentioning

confidence: 99%

“…Although such observations are suggestive of a role for cell-cell interactions in mediating IIF in tissue, these studies have introduced a host of confounding factors by comparing freshly trypsinized cells with cells cultured for various lengths of time in different configurations. To whit, the probability of IIF is known to be affected both by trypsinization (Sandler and Andersson, 1984;Yarmush et al, 1992) and time in culture (Hetzel et al, 1973;Hornung et al, 1996;Darr and Hubel, 2001); in cell clusters, mass transport limitations may also affect IIF (Levin et al, 1977).…”

Section: Introductionmentioning

confidence: 99%

Kinetics and Mechanism of Intercellular Ice Propagation in a Micropatterned Tissue Construct

Irimia

Karlsson

2002

Biophysical Journal

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Understanding the effects of cell-cell interaction on intracellular ice formation (IIF) is required to design optimized protocols for cryopreservation of tissue. To determine the effects of cell-cell interactions during tissue freezing, without confounding effects from uncontrolled factors (such as time in culture, cell geometry, and cell-substrate interactions), HepG2 cells were cultured in pairs on glass coverslips micropatterned with polyethylene glycol disilane, such that each cell interacted with exactly one adjacent cell. Assuming the cell pair to be a finite state system, being either in an unfrozen state (no ice in either cell), a singlet state (IIF in one cell only), or a doublet state (IIF in both cells), the kinetics of state transitions were theoretically modeled and cryomicroscopically measured. The rate of intercellular ice propagation, estimated from the measured singlet state probability, increased in the first 24 h of culture and remained steady thereafter. In cell pairs cultured for 24 h and treated with the gap junction blocker 18beta-glycyrrhetinic acid before freezing, the intercellular ice propagation rate was lower than in untreated controls (p < 0.001), but significantly greater than zero (p < 0.0001). These results suggest that gap junctions mediate some, but not all, mechanisms of ice propagation in tissue.

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“…Different cryopreservation methodologies alternative to the commonly applied protocols are reported in the literature; some refer to the use of structured substrates, 15 cell 1 Animal Cell Technology, IBET=ITQB-UNL, Oeiras, Portugal. 2 Kryobiophysik & Kryotechnologie, Fraunhofer-IBMT, Ingbert, Germany.…”

Section: Introductionmentioning

confidence: 99%

Cryopreservation of Adherent Cells: Strategies to Improve Cell Viability and Function After Thawing

Malpique¹,

Ehrhart

Katsen-Globa

et al. 2009

Tissue Engineering Part C: Methods

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The commonly applied cryopreservation protocols routinely used in laboratories worldwide were developed for simple cell suspensions, and their application to complex systems, such as cell monolayers, tissues, or biosynthetic constructs, is not straightforward. In particular for monolayer cultures, cell detachment and membrane damage are often observed after cryopreservation. In this work, combined strategies for the cryopreservation of cells attached to Matrigel-coated well plate's surfaces were investigated based on cell entrapment in clinicalgrade, ultra-high viscosity alginate using two cell lines, neuroblastoma N2a and colon adenocarcinoma Caco-2, with distinct structural and functional characteristics. As the cryopreservation medium, serum-free CryoStor solution was compared with serum-supplemented culture medium, both containing 10% DMSO. Using culture medium, entrapment beneath an alginate layer was needed to improve cell recovery by minimizing membrane damage and cell detachment after thawing; nevertheless, up to 50% cell death still occurred within 24 h after thawing. The use of CryoStor solution represented a considerable improvement of the cryopreservation process for both cell lines, allowing the maintenance of high postthaw membrane integrity as well as full recovery of metabolic activity and differentiation capacity within 24 h postthawing; in this case, entrapment beneath an alginate layer did not confer further protection to cryopreserved Caco-2 cells, but was crucial for maintenance of attachment and integrity of N2a neuronal networks.

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Cryopreservation of Anchorage-Dependent Mammalian Cells Fixed to Structured Glass and Silicon Substrates

Cited by 29 publications

References 0 publications

Cryopreservation of Fibroblasts Immobilized Within a Porous Scaffold: Effects of Preculture and Collagen Coating of Scaffold on Performance of Three‐Dimensional Cryopreservation

Cryopreservation of Fibroblasts Immobilized Within a Porous Scaffold: Effects of Preculture and Collagen Coating of Scaffold on Performance of Three‐Dimensional Cryopreservation

Kinetics and Mechanism of Intercellular Ice Propagation in a Micropatterned Tissue Construct

Cryopreservation of Adherent Cells: Strategies to Improve Cell Viability and Function After Thawing

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