Bovine and porcine fibroblasts can be immortalized with intact karyotype by the expression of mutant cyclin dependent kinase 4, cyclin D, and telomerase

Donai, Kenichiro; Kiyono, Tohru; Eitsuka, Takahiro; Guo, Yi; Kuroda, Kensuke; Sone, Hirohito; Isogai, Emiko; Fukuda, T.

doi:10.1016/j.jbiotec.2014.02.017

Cited by 43 publications

(43 citation statements)

References 36 publications

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“…In the immortalized cells, the CDK4 R24C mutant does not bind to p16 and thus avoids negative regulation. The amino acid sequences of CDK4 and cyclin D are evolutionally well conserved, meaning that this immortalization method could potentially be applied to various other species, including pigs, cattle, and monkeys in addition to prairie voles [6,7,8]. Furthermore, the immortalized prairie vole cells retained both the chromosomal pattern and differentiation ability of the primary cells from which they were derived.…”

Section: Discussionmentioning

confidence: 99%

“…The accumulation of p16 is a recognized molecular mechanism for the development of cellular senescence. Further, we reported previously that the combined expression of mutated cyclin-dependent kinase 4 (CDK4), cyclin D, and the enzymatic subunit of telomerase reverse transcriptase (TERT) allows the cells of multiple species, including cattle, swine, monkeys, and prairie voles, to bypass cellular senescence [6,7,8]. The amino acid sequences of CDK4 and cyclin D are evolutionally well conserved across organisms, and thus, the expression of mutated versions of these genes allows senescence signals to be bypassed in the cells of various species.…”

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confidence: 99%

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Immortalized prairie vole-derived fibroblasts (VMF-K4DTs) can be transformed into pluripotent stem cells and provide a useful tool with which to determine optimal reprogramming conditions

Katayama

Hirayama

Kiyono

et al. 2017

Journal of Reproduction and Development

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The cellular conditions required to establish induced pluripotent stem cells (iPSCs), such as the number of reprogramming factors and/or promoter selection, differ among species. The establishment of iPSCs derived from cells of previously unstudied species therefore requires the extensive optimization of programming conditions, including promoter selection and the optimal number of reprogramming factors, through a trial-and-error approach. While the four Yamanaka factors Oct3/4, Sox2, Klf4, and c-Myc are sufficient for iPSC establishment in mice, we reported previously that six reprogramming factors were necessary for the creation of iPSCs from primary prairie vole-derived cells. Further to this study, we now show detailed data describing the optimization protocol we developed in order to obtain iPSCs from immortalized prairie vole-derived fibroblasts. Immortalized cells can be very useful tools in the optimization of cellular reprogramming conditions, as cellular senescence is known to dramatically decrease the efficiency of iPSC establishment. The immortalized prairie vole cells used in this optimization were designated K4DT cells as they contained mutant forms of CDK4, cyclin D, and telomerase reverse transcriptase (TERT). We show that iPSCs derived from these immortalized cells exhibit the transcriptional silencing of exogenous reprogramming factors while maintaining pluripotent cell morphology. There were no observed differences between the iPSCs derived from primary and immortalized prairie vole fibroblasts. Our data suggest that cells that are immortalized with mutant CDK4, cyclin D, and TERT provide a useful tool for the determination of the optimal conditions for iPSC establishment.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Immortalized prairie vole-derived fibroblasts (VMF-K4DTs) can be transformed into pluripotent stem cells and provide a useful tool with which to determine optimal reprogramming conditions

Katayama

Hirayama

Kiyono

et al. 2017

Journal of Reproduction and Development

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“…The BFF_NCC1 cells were kindly provided by Prof. Fukuda (17). Cells were cultured in DMEM culture medium (Nacalai-tesque, Japan) including 10% fetal bovine serum (FBS; Biowest SAS, France) and 1% antibiotic and antimycotic solution (Nacalai-tesque, Japan).…”

Section: Methodsmentioning

confidence: 99%

Inhibitory effect of BMAP-28 on Leptospiral Lipopolysaccharide-Induced TLR2-Dependent Immune Response in Bovine Cells

Guo

Zhang

et al. 2016

Jundishapur J Microbiol

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BackgroundBovine leptospirosis is a widespread zoonotic disease, leading to serious economic losses in animal production and causing potential hazards to human health. Leptospiral lipopolysaccharide (L-LPS) plays an important role in leptospirosis pathogenicity.ObjectivesWith respect to L-LPS endotoxin-like activity, we examined bovine immune response to L-LPS and the inhibitory ability of bovine myeloid antimicrobial peptide-28 (BMAP-28) against L-LPS-induced immune activation in bovine cells.Materials and MethodsIn this study, L-LPS-induced proinflammatory cytokine production in bovine cells was quantitatively measured with real-time PCR and ELISA, and we determined which cell membrane receptors (toll-like receptor [TLR]2 and TLR4) played a major role. In addition, the ability of BMAP-28 to inhibit L-LPS-induced endotoxin-like immune activation in bovine cells was determined by the decrease in cytokine secretion.ResultsL-LPS showed the ability to induce cytokine production in bovine cells, and its induction was TLR2-dependent. BMAP-28 was used to inhibit L-LPS-induced endotoxin-like activity. The function of BMAP-28 was to inhibit LPS-induced TLR2 expression and cytokine production.ConclusionsIn this study, the L-LPS immune response of bovine cells was significant, indicating that TLR2 is the predominant receptor for L-LPS. Due to L-LPS endotoxin-like activity, we found a strategy through using BMAP-28 to prevent L-LPS-induced TLR2-dependent immune activation in bovine cells.

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“…Our group reported earlier that expression of mutant CDK4 (R24C), Cyclin D, and TERT allows us to induce the infinite cell proliferation (immortalization) of the bovine and porcine derived cells. These cells are named K4DT (CDK4-Cyclin D-TERT) cells (Donai et al 2014;Kuroda et al 2015a, b;Shiomi et al 2011) from their name of the introduced genes. Since the K4DT cell lines infinitely proliferate with keeping the original nature of the cells (Shiomi et al 2011).…”

Section: Introductionmentioning

confidence: 99%

Cellular conservation of endangered midget buffalo (Lowland Anoa, Bubalus quarlesi) by establishment of primary cultured cell, and its immortalization with expression of cell cycle regulators

et al. 2016

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Lowland Anoa has become endangered due to hunting and human activity. Protection and breeding of endangered species in a controlled environment is the best way of conservation. However, it is not possible to adopt this approach for all endangered species because of the cost involved and the everincreasing number of critically endangered species. In consideration of these limitations to the conventional conservation methods, we established a primary cell culture of endangered buffalo (Lowland Anoa, Bubalus quarlesi), for the preservation of this biological resource. In addition, we introduced human derived, mutant cyclin dependent kinase 4 (CDK4), Cyclin D, and telomerase reverse transcriptase (TERT) into the primary cells. The successful introduction of these three genes was confirmed by western blot with specific antibodies, and enzymatic activity. 123 Cytotechnology (2016) 68:1937-1947 DOI 10.1007 We also showed that the expression of mutant CDK4, Cyclin D, and TERT allows us to efficiently establish an immortalized cell line, with an intact chromosome pattern, from Lowland Anoa. To the best of our knowledge, this study is the first investigation that established an immortalized cell line of an endangered wild animal species.

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Bovine and porcine fibroblasts can be immortalized with intact karyotype by the expression of mutant cyclin dependent kinase 4, cyclin D, and telomerase

Cited by 43 publications

References 36 publications

Immortalized prairie vole-derived fibroblasts (VMF-K4DTs) can be transformed into pluripotent stem cells and provide a useful tool with which to determine optimal reprogramming conditions

Immortalized prairie vole-derived fibroblasts (VMF-K4DTs) can be transformed into pluripotent stem cells and provide a useful tool with which to determine optimal reprogramming conditions

Inhibitory effect of BMAP-28 on Leptospiral Lipopolysaccharide-Induced TLR2-Dependent Immune Response in Bovine Cells

Cellular conservation of endangered midget buffalo (Lowland Anoa, Bubalus quarlesi) by establishment of primary cultured cell, and its immortalization with expression of cell cycle regulators

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