2021
DOI: 10.3390/ani11020367
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Establishing Cell Lines from Fresh or Cryopreserved Tissue from the Great Crested Newt (Triturus cristatus): A Preliminary Protocol

Abstract: This study describes a successful protocol for establishing cell lines from the threatened Triturus cristatus in terms of collection, preparing, establishing, cryopreserving, thawing and quality checking. Different parameters such as media, media change, fresh vs. cryopreserved tissue and seeding density were tested to optimize culture conditions for this species. With fresh tissue, no considerable differences in the use of two different media were found, but with cryopreserved tissue, a combination of ITS (in… Show more

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Cited by 5 publications
(11 citation statements)
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“…Compared to previous reports (Ferretti & Brockes, 1988;Strand et al, 2021), our procedure to grow primary newt tissue cells is better as we achieved contamination-free explant culture with 100% attachment of explants on both coated and non-coated surfaces.…”
Section: Discussionmentioning
confidence: 82%
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“…Compared to previous reports (Ferretti & Brockes, 1988;Strand et al, 2021), our procedure to grow primary newt tissue cells is better as we achieved contamination-free explant culture with 100% attachment of explants on both coated and non-coated surfaces.…”
Section: Discussionmentioning
confidence: 82%
“…However, 5% CO 2 has been supplied in the case of T. cristatus (Strand et al, 2021), and a temperature of 24 C has been reported for growing cells from limb tissue of N. viridescens (Ferretti & Brockes, 1988) and a temperature of 28 C has been reported for culturing cells of T. cristatus (Strand et al, 2021). Perhaps, Strand et al (2021) used 5%…”
Section: Discussionmentioning
confidence: 99%
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“…One main challenge with amphibian skin cryopreservation and cell culture is contamination from bacteria and fungi ( Strauß et al 2013 ). Strand et al (2021) have shown that even with extensive washing, contamination can still be problematic, especially as liquid nitrogen is known to not fully inhibit the replication of microorganisms ( Bajerski et al 2020 ). Though much work is yet to be done, amphibian cryobanking, ART and aART are undeniably exciting and hold great promise as conservation safety nets.…”
Section: Introductionmentioning
confidence: 99%
“…However, the San Diego Zoo Institute for Conservation Research Frozen Zoo® already holds a large collection of cryopreserved amphibian tissue and cell lines(Chemnick et al, 2009) and the IUCN amphibian specialist group have biobank and ART working groups, so advances are being made in this area. One main challenge with amphibian skin cryopreservation and cell culture is contamination from bacteria and fungi(Strauß et al, 2013) Strand et al, 2021. have shown that even with extensive washing, contamination can still be problematic, especially as liquid nitrogen is known to not fully inhibit the replication of microorganisms(Bajersk et al, 2020).…”
mentioning
confidence: 99%