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

Fukuda, T.; Iino, Y.; Eitsuka, Takahiro; Onuma, Manabu; Katayama, Masafumi; Murata, Koichi; Inoue‐Murayama, Miho; Hara, Koji; Isogai, Emiko; Kiyono, Tohru

doi:10.1007/s10616-016-0004-0

Cited by 26 publications

(21 citation statements)

References 12 publications

(13 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Our research group previously reported that combined expression of R24C mutant cyclin-dependent kinase 4 (CDK4), Cyclin D1, and telomere reverse transcriptase (TERT) allowed us to efficiently immortalize human- (Shiomi et al, 2011), cattle and pig- (Donai et al, 2014), prairie vole- (Katayama et al, 2016(Katayama et al, , 2017, monkey- (Kuroda et al, 2015a), midget buffalo- (Fukuda et al, 2016), and mega bat- (Tani et al, 2019), Tsushima wildcat-derived cells (Gouko et al, 2018). Furthermore, growth acceleration with mutant CDK4 and Cyclin D1 is conserved even in sea turtles, suggesting that the underlying cell cycle mechanism was well-conserved throughout animal evolution (Fukuda et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Human Derived Immortalized Dermal Papilla Cells With a Constant Expression of Testosterone Receptor

Fukuda

Takahashi

Takase

et al. 2020

Front. Cell Dev. Biol.

Self Cite

View full text Add to dashboard Cite

Androgenetic alopecia (AGA) is the most common type of hair loss, and is mainly caused by the biological effects of testosterone on dermal papilla cells (DPCs). In vitro culturing of DPCs might be a useful tool for the screening of target molecule of AGA. However, primary DPCs cannot continuously proliferate owing to cellular senescence and cell culture stress. In this study, we introduced mutant cyclin-dependent kinase 4 (CDK4), Cyclin D1, and telomerase reverse transcriptase (TERT) into DPCs. We confirmed protein expression of CDK4 and Cyclin D1, and enzymatic activity of TERT. Furthermore, we found the established cell line was free from cellular senescence. We also introduced the androgen receptor gene using a recombinant retrovirus, to compensate the transcriptional suppressed endogenous androgen receptor in the process of cell proliferation. Furthermore, we detected the efficient nuclear translocation of androgen receptor into the nucleus after the treatment of dihydrotestosterone, indicating the functionality of our introduced receptor. Our established cell line is a useful tool to identify the downstream signaling pathway, which activated by the testosterone.

show abstract

Section: Introductionmentioning

confidence: 99%

Human Derived Immortalized Dermal Papilla Cells With a Constant Expression of Testosterone Receptor

Fukuda

Takahashi

Takase

et al. 2020

Front. Cell Dev. Biol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Our research group has previously reported efficient immortalization in multiple species via co-expression of R24C mutant cyclin-dependent kinase 4 (CDK4 R24C ), Cyclin D1, and telomere reverse transcriptase (TERT) [8][9][10][11][12][13][14]. This immortalization method using mutant CDK4, Cyclin D1, and TERT was termed "K4DT" in reference to the introduced genes.…”

Section: Introductionmentioning

confidence: 99%

Efficient immortalization of human dental pulp stem cells with expression of cell cycle regulators with the intact chromosomal condition

et al. 2020

Self Cite

View full text Add to dashboard Cite

Clinical studies have recently demonstrated that autologous transplantation of mobilized dental pulp stem cells is a safe and efficacious potential therapy for pulp regeneration. However, some limitations need to be addressed, such as the high cost of the safety and quality control tests for isolated individual dental pulp cell products before transplantation. Therefore, more efficient in vitro culturing of human dental pulp stem cells might be useful for providing low cost and high reliability testing for pulp regeneration therapy. In this study, we established a novel immortalized dental pulp stem cell line by co-expressing a mutant cyclin-dependent kinase 4 (CDK4 R24C), Cyclin D1, and telomerase reverse transcriptase (TERT). The established cell line maintained its original diploid chromosomes and stemness characteristics and exhibited an enhanced proliferation rate. In addition, we showed the immortalized human dental pulp stem cells still keeps their osteogenic and adipogenic differentiation abilities under appropriate culture conditions even though the cell proliferation was accelerated. Taken together, our established cell lines could serve as a useful in vitro tool for pulp regeneration therapy, and can contribute to reproducibility and ease of cell handling, thereby saving time and costs associated with safety and quality control tests.

show abstract

“…Furthermore, our research group previously reported that expression of human-derived mutant Cyclin-dependent kinase 4 (CDK4), Cyclin D and Telomerase Reverse Transcriptase (TERT) efficiently immortalises cells of various mammalian species, including human, bovine 8 , swine 9 , 10 , monkey 11 , prairie vole 12 , 13 and midget buffalo 14 . We designated this recently developed method as K4DT, based on the identities of the introduced genes (mutant CD K4 , Cyclin D and T ERT).…”

Section: Introductionmentioning

confidence: 99%

Expression of human mutant cyclin dependent kinase 4, Cyclin D and telomerase extends the life span but does not immortalize fibroblasts derived from loggerhead sea turtle (Caretta caretta)

Fukuda

Eitsuka

Donai

et al. 2018

Sci Rep

Self Cite

View full text Add to dashboard Cite

Conservation of the genetic resources of endangered animals is crucial for future generations. The loggerhead sea turtle (Caretta caretta) is a critically endangered species, because of human hunting, hybridisation with other sea turtle species, and infectious diseases. In the present study, we established primary fibroblast cell lines from the loggerhead sea turtle, and showed its species specific chromosome number is 2n = 56, which is identical to that of the hawksbill and olive ridley sea turtles. We first showed that intensive hybridization among multiple sea turtle species caused due to the identical chromosome number, which allows existence of stable hybridization among the multiple sea turtle species. Expressions of human-derived mutant Cyclin-dependent kinase 4 (CDK4) and Cyclin D dramatically extended the cell culture period, when it was compared with the cell culture period of wild type cells. The recombinant fibroblast cell lines maintained the normal chromosome condition and morphology, indicating that, at the G1/S phase, the machinery to control the cellular proliferation is evolutionally conserved among various vertebrates. To our knowledge, this study is the first to demonstrate the functional conservation to overcome the negative feedback system to limit the turn over of the cell cycle between mammalian and reptiles. Our cell culture method will enable the sharing of cells from critically endangered animals as research materials.

show abstract

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

Cited by 26 publications

References 12 publications

Human Derived Immortalized Dermal Papilla Cells With a Constant Expression of Testosterone Receptor

Human Derived Immortalized Dermal Papilla Cells With a Constant Expression of Testosterone Receptor

Efficient immortalization of human dental pulp stem cells with expression of cell cycle regulators with the intact chromosomal condition

Expression of human mutant cyclin dependent kinase 4, Cyclin D and telomerase extends the life span but does not immortalize fibroblasts derived from loggerhead sea turtle (Caretta caretta)

Contact Info

Product

Resources

About