Establishment of immortalized primary cell from the critically endangered Bonin flying fox (Pteropus pselaphon).

Tetsuya Tani,Hajime Suzuki,Takashi Nagamine,Tohru Kiyono,Tomokazu Fukuda,Manabu Onuma,Miho Inoue-Murayama,Masafumi Katayama,Takahiro Eitsuka,Kiyotaka Nakagawa,Yumiko Nakaya,Zhilong Yang

doi:10.1371/journal.pone.0221364

Abstract

The Bonin flying fox (Pteropus pselaphon) is one of the most critically endangered species of animals. The number of this species is estimated to be around 150; being classified at the top rank in the list by International Union of Animal Conservation. Our group previously showed that expression of CDK4, CYCLIN D1, and telomerase reverse transcriptase (TERT) efficiently induce immortalization of human, bovine, swine, monkey, and buffalo-derived cells. In this manuscript, we successfully established the primary cells from Bonin flying fox. We introduced CDK4, CYCLIN D1, and TERT into the primary cells. The established cells showed efficient expression of introduced genes at the protein level. Furthermore, the established cells were free from senescence, indicating it reached to immortalization. Moreover, we showed that interspecies somatic cell nuclear transfer of Bonin flying fox derived cell into bovine embryo allowed the development of the embryo to 8 cell stages. Our established cell has the potential to contribute to species conservation.

Highlights

The Bonin flying fox (Pteropus pselaphon) is an original species in North and South Iwo Jima, the regions of Bonin Islands of Japan
We showed that the expressions of mutant cyclin dependent kinase (CDK4) and CYCLIN D1, and telomerase reverse transcriptase (TERT) induce the immortalization in multiple species [1,2,3,4,5,6,7]
We introduced mutant cyclin dependent kinase 4 (CDK4), CYCLIN D1, and TERT as a mixture

Summary

Introduction

The Bonin flying fox (Pteropus pselaphon) is an original species in North and South Iwo Jima, the regions of Bonin Islands of Japan. During World War II, Bonin flying fox was hunted and exported as military food to Guam Island. Due to the hunting practices for food purpose, and forest destruction by human activity, the number of Bonin flying fox species significant decreased around the early 1970s. Bonin flying fox was believed to have reached extinction, since there was no report on the evidence of its survival. In 1986, a colony of Bonin flying fox was found in a cave of Chichijima, one of the Bonin Islands.

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