Viability of Cell Cultures Following Extended Preservation in Liquid Nitrogen

Greene, A.E.; Athreya, Balu H.; Lehr, Herndon B.; Coriell, Lewis L.

doi:10.3181/00379727-124-31992

Cited by 15 publications

(4 citation statements)

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“…1a) forming primmorphs in the usual 2-day period (Fig 1c). The same dissociated cells, when subjected to the freezing and thawing procedure, gave an amount of living cells consistent with literature values for mammalian and insect cells (Green et al 1967;Freshney 1994). Notwithstanding this, the standard horizontal agitation, usually leading to the formation of primmorphs from unfrozen cells, did not produce any, but only promoted simple sponge cells re-aggregation not organized in primmorphs (Fig.…”

Section: Discussionsupporting

confidence: 86%

Primmorphs Cryopreservation: A New Method for Long-Time Storage of Sponge Cells

et al. 2012

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The possibility to cryopreserve cells allows for wide opportunities of flexible handling of cell cultures from different sponge species. Primmorphs model, a multicellular 3D aggregate formed by dissociated sponge cells, is considered one of the best approaches to establish sponge cell culture but, in spite of the available protocols for freezing sponge cells, there is no information regarding the ability of the latter to form primmorphs after thawing. In the present work, we demonstrate that, after a freezing and thawing cycle using dissociated Petrosia ficiformis cells as a model, cells viability was high but it was not possible to obtain primmorphs. The same protocol for cryopreservation was then used to directly freeze primmorphs. In this second case, after thawing, viability and the cellular proliferative level were similar to unfrozen standard primmorphs. Spiculogenesis in thawed primmorphs was evaluated by quantifying the silicatein gene expression level and by assaying the silica amount in the newly formed spicules, then compared with the correspondent values obtained in standard unfrozen primmorphs. Results indicate that the freezing cycle does not affect the spiculogenesis rate. Finally, the expression level of heat shock protein 70, a well-known stress marker, was assayed and the results showed no differences between frozen and unfrozen samples. These findings are likely to promote relevant improvements in sponge cell culture technique, allowing for a worldwide exchange of living biological material, paving the way for cell banking of Porifera.

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Section: Discussionsupporting

confidence: 86%

Primmorphs Cryopreservation: A New Method for Long-Time Storage of Sponge Cells

et al. 2012

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“…Therefore, the cryopreservation of viable somatic cell tissues is quickly assuming an increasingly important role in BRBs (Ryder 2002). Although these tissue samples are also finite, they can be propagated using specific culture conditions before being cryopreserved and stored for many years (Green 1967).…”

Section: Introductionmentioning

confidence: 99%

Isolation, culture and characterisation of somatic cells derived from semen and milk of endangered sheep and eland antelope

Nel-Themaat

Gómez

Damiani

et al. 2007

Reprod. Fertil. Dev.

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Semen and milk are potential sources of somatic cells for genome banks. In the present study, we cultured and characterised cells from: (1) cooled sheep milk; (2) fresh, cooled and frozen-thawed semen from Gulf Coast native (GCN) sheep (Ovis aries); and (3) fresh eland (Taurotragus oryx) semen. Cells attached to the culture surface from fresh (29%), cooled (43%) and slow-frozen (1 degrees C/min; 14%) ram semen, whereas no attachment occurred in the fast-frozen (10 degrees C/min) group. Proliferation occurred in fresh (50%) and cooled (100%) groups, but no cells proliferated after passage 1 (P1). Eland semen yielded cell lines (100%) that were cryopreserved at P1. In samples from GCN and cross-bred milk, cell attachment (83% and 95%, respectively) and proliferation (60% and 37%, respectively) were observed. Immunocytochemical detection of cytokeratin indicated an epithelial origin of semen-derived cells, whereas milk yielded either fibroblasts, epithelial or a mixture of cell types. Deoxyribonucleic acid microsatellite analysis using cattle-derived markers confirmed that eland cells were from the semen donor. Eland epithelial cells were transferred into eland oocytes and 12 (71%), six (35%) and two (12%) embryos cleaved and developed to morulae or blastocyst stages, respectively. In conclusion, we have developed a technique for obtaining somatic cells from semen. We have also demonstrated that semen-derived cells can serve as karyoplast donors for nuclear transfer.

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“…This beneficial effect of serum might be related to avoidance of osmotic shock during thawing of cells. Studies in the late 1960s indicated that cryopreserved cells in medium with serum and anti-freeze agent were viable after 4.5 years of storage [11]. Other fundamental issues noted early on were the superior effect of slow cooling rate (approximately -1C/min) and fast warming rate for the viability of the cryopreserved cells [12;13].…”

Section: Principles Of Cryopreservation Of Cellsmentioning

confidence: 99%

Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies

et al. 2021

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DNA damage and repair activity are often assessed in blood s#38les from humans in different types of molecular epidemiology studies. However, it is not always feasible to analyse the s#38les on the day of collection without any type of storage. For instance, certain studies use repeated s#38ling of cells from the same subject or s#38les from different subjects collected at different time-points, and it is desirable to analyse all these s#38les in the same comet assay experiment. In addition, flawless comet assay analyses on frozen s#38les opens up for the possibility of using this technique on biobank material. In this article we discuss the use of cryopreserved peripheral blood mononuclear cells (PBMCs), buffy coat (BC) and whole blood (WB) for analysis of DNA damage and repair using the comet assay. The published literature and the authors’ experiences indicate that various types of blood s#38les can be cryopreserved with only minor effect on the basal level of DNA damage. There is evidence to suggest that WB and PBMCs can be cryopreserved for several years without much effect on the level of DNA damage. However, care should be taken when cryopreserving WB and BCs. It is possible to use either fresh or frozen s#38les of blood cells, but results from fresh and frozen cells should not be used in the same dataset. The article outlines detailed protocols for the cryopreservation of PBMCs, BCs and WB s#38les.

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Viability of Cell Cultures Following Extended Preservation in Liquid Nitrogen

Cited by 15 publications

References 0 publications

Primmorphs Cryopreservation: A New Method for Long-Time Storage of Sponge Cells

Primmorphs Cryopreservation: A New Method for Long-Time Storage of Sponge Cells

Isolation, culture and characterisation of somatic cells derived from semen and milk of endangered sheep and eland antelope

Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies

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