A protocol for the cryoconservation of breeds by low-cost emergency cell banks – a pilot study

Groeneveld, Eildert; Tinh, Nguyen Huu; Kues, Wilfried A.; Vien, Nguyen Thi

doi:10.1017/s1751731107000869

Cited by 20 publications

(15 citation statements)

References 17 publications

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“…To gain a better understanding of shrimp growth and the underlying molecular mechanisms, we initiate a research project to identify muscle structural and regulatory genes by cDNA library and gene expression analysis. In a previous study, the abdominal muscle cDNA library of shrimp L.vannamei was successfully constructed [6]. Preliminary data analysis suggests that abundant and diverse transcripts are present in the cDNA library established by our laboratory.…”

Section: Introductionmentioning

confidence: 90%

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Molecular Characterizations of a Novel Putative DNA-Binding Protein LvDBP23 in Marine Shrimp L. vannamei Tissues and Molting Stages

et al. 2011

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Background Litopenaeus Vannamei, well known as pacific white shrimp, is the most popular shrimp in the world shrimp market. Identification and characterization of shrimp muscle regulatory genes are not only important for shrimp genetic improvement, but also facilitate comparative genomic tools for understanding of muscle development and regeneration.Methodology/Principal FindingsA novel mRNA encoding for a putative DNA-binding protein LvDBP23 was identified from Litopenaeus vannamei abdominal muscle cDNA library. The LvDBP23 cDNA contains 639 nucleotides of protein-coding sequence with deduced 212 amino acids of predicted molecular mass 23.32 kDa with glycine-rich domain at amino acid position 94–130. The mRNA sequence is successfully used for producing LvDBP23 recombinant protein in sf9 insect cell expression system. The expression of LvDBP23 mRNA is presented in abdominal muscle and swimming leg muscle, as well as other tissues including intestine, lymphoid and gill. The mRNA expression has the highest level in abdominal muscle in all tested tissues. LVDBP23 transcript during the molt cycle is highly expressed in the intermolt stage. In vitro nucleic acid-binding assays reveal that LvDBP23 protein can bind to both ssDNA and dsDNA, indicating its possible role of regulation of gene transcription.Conclusions/SignificanceWe are the first to report a DNA-binding protein identified from the abdominal muscle tissue of marine shrimp L. Vannamei. Its high-level specific expression during the intermot stage suggests its role in the regulation of muscle buildup during the growth phase of shrimp molt cycle.

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Section: Introductionmentioning

confidence: 90%

“…Previously, we have described the L. vannamei muscle cDNA library [6]. Positive muscle cDNA clones were identified by using phage lift hybridization method as described by [11]).…”

Section: Methodsmentioning

confidence: 99%

Molecular Characterizations of a Novel Putative DNA-Binding Protein LvDBP23 in Marine Shrimp L. vannamei Tissues and Molting Stages

et al. 2011

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Section: Introductionunclassified

“…Contudo, muitas dessas ações são inviáveis para a maioria das nações menos desenvolvidas (1) . Assim, bancos genéticos baseados em células somáticas poderiam universalizar ações preservacionistas de maneira fácil e com custos bastante reduzidos (1) . A metodologia permitiria transpor limitações produzidas por animais incapacitados de produzir gametas (castração, freemartinismo), ou em epizootias dramáticas que determinem o abate sanitário dos animais (2) .…”

Section: Introductionunclassified

Células Fetais Bovinas De Cultivo Primário Submetidas a Diferentes Pressões Negativas Antes Do Congelamento Em Palhetas

et al. 2018

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ResumoO congelamento de células é uma importante ferramenta na preservação de espécies ameaçadas de extinção. Células fetais de cultivo primário obtidas de um bovino clone foram submetidas à pressão negativa (PN) de 200, 500 ou 800 mbar, imediatamente (PN0h) ou três horas antes (PN3h) do congelamento em palhetas finas, com 10% de DMSO como crioprotetor. Células frescas e congeladas sem submissão à PN foram utilizadas como controles. Avaliou-se a viabilidade pós-descongelamento, a curva de proliferação celular, assim como o tempo de duplicação da população (PDT) celular, a cada 24 horas, durante oito dias. Os dados obtidos foram submetidos ao teste de Tukey ou Qui quadrado (P≤0,05). A sobrevivência média dos grupos controle (89,8%) e PN500 0h (88,1%) foi superior aos outros grupos; o tempo de PDT foi semelhante nos grupos fresco (27,5 ± 0,35 h), controle congelado (30,1 ± 2,3 h) e PN500 0h (32,4 ± 1,6 h). O menor tempo foi observado no grupo PN800 0h (21,9 h). O congelamento de células fetais bovinas de cultivo primário, realizado em palhetas de 0,25 mL, com 10% de DMSO, possibilita elevadas taxas de sobrevivência após o descongelamento. A PN modifica a curva de crescimento de células criopreservadas, sendo que as intensidades de 200 ou 500 mbar, aplicadas imediatamente antes do congelamento das células, possibilitam curvas de proliferação semelhantes às obtidas com células frescas. Palavras-chave: células somáticas; criopreservação; estresse controlado; preservação animal. AbstractCell cryopreservation is an important tool in the preservation of endangered species. Fetal bovine primary culture-obtained cells were subjected to negative pressure (NP) 200, 500 or 800 mbar, just before (NP0h) or 3 hours before (NP3h) freezing into fine (0.25 mL) straws, using 10% DMSO as cryoprotectant. Fresh and frozen fibroblasts non submitted to NP were used as controls. We evaluated cell viability after freezing, cell proliferation curve, and population doubling time (PDT) every 24

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“…Reports on cryobanking costs are scarce, despite costs being a limiting factor in the establishment of gene banks (McClintock et al 2007;Groeneveld et al 2008), or only based on simulations (Gandini et al 2007). Information about cryoconservation costs is needed in order to optimise conservation programmes.…”

Section: Introductionmentioning

confidence: 99%

Implementation and cost analysis of a regional farm animal cryobank: an Italian case study

Pizzi

Turri

Gliozzi

et al. 2016

Italian Journal of Animal Science

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Scientific and technical reports on farm animal genetic resources (AnGR) cryoconservation activities, and related costs, are needed to optimise conservation programmes. In this paper, we presented the recent Italian AnGR gene banking development, including the creation of the first regional animal cryobank, the 'Lombardia Farm Animal Genetic Resources Cryobank' (LABank). In order to provide indications on cryobanking costs, a detailed analysis of the expenses incurred during the creation of LABank was carried out. Currently, in LABank genetic material (spermatozoa, blood and hair bulbs) of Italian cattle, sheep and goat local breeds is cryopreserved, for a total amount of approximately 2500 semen doses collected from 46 donors of five local breeds. The costs incurred by creating the semen storage showed differences among species and semen collection procedures, providing indications to enhance the setting up of regional cryobanks. ARTICLE HISTORY

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A protocol for the cryoconservation of breeds by low-cost emergency cell banks – a pilot study

Cited by 20 publications

References 17 publications

Molecular Characterizations of a Novel Putative DNA-Binding Protein LvDBP23 in Marine Shrimp L. vannamei Tissues and Molting Stages

Molecular Characterizations of a Novel Putative DNA-Binding Protein LvDBP23 in Marine Shrimp L. vannamei Tissues and Molting Stages

Células Fetais Bovinas De Cultivo Primário Submetidas a Diferentes Pressões Negativas Antes Do Congelamento Em Palhetas

Implementation and cost analysis of a regional farm animal cryobank: an Italian case study

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