title: 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.
abstract: 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 ever-increasing 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. 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.
authors.affiliation: United Graduate School of Agricultural Sciences, Graduate School of Agricultural Science, Iwate University, 4-3-5, Ueda, Morioka, Iwate, 020-8551, Japan. tomofukuda009@gmail.com.|Wildlife Genome Collaborative Research Group, National Institute for Environmental Studies, Tsukuba, Ibaraki, Japan. tomofukuda009@gmail.com.
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Fukuda
authors.affiliation: Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, Japan.
authors.forename: Yuuka
authors.initials: Y
authors.lastname: Iino
authors.affiliation: Faculty of Applied Life Sciences, Niigata University of Pharmacy and Applied Life Sciences, Niigata, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Eitsuka
authors.affiliation: Wildlife Genome Collaborative Research Group, National Institute for Environmental Studies, Tsukuba, Ibaraki, Japan.|Ecological Risk Assessment and Control Section, Center for Environmental Biology and Ecosystem, National Institute for Environmental Studies, Tsukuba, Ibaraki, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Onuma
authors.affiliation: Wildlife Genome Collaborative Research Group, National Institute for Environmental Studies, Tsukuba, Ibaraki, Japan.|Ecological Risk Assessment and Control Section, Center for Environmental Biology and Ecosystem, National Institute for Environmental Studies, Tsukuba, Ibaraki, Japan.
authors.forename: Masafumi
authors.initials: M
authors.lastname: Katayama
authors.affiliation: Laboratory of Wildlife Science, College of Bio-resource Sciences, Nihon University, Fujisawa, Kanagawa, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Murata
authors.affiliation: Wildlife Genome Collaborative Research Group, National Institute for Environmental Studies, Tsukuba, Ibaraki, Japan.|Wildlife Research Center, Kyoto University, Kyoto, Japan.
authors.forename: Miho
authors.initials: M
authors.lastname: Inoue-Murayama
authors.affiliation: Kanazawa Zoological Gardens, Yokohama Greenery Foundation, Yokohama, Kanagawa, Japan.
authors.forename: Kumiko
authors.initials: K
authors.lastname: Hara
authors.affiliation: Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, Japan.
authors.forename: Emiko
authors.initials: E
authors.lastname: Isogai
authors.affiliation: Division of Carcinogenesis and Prevention, National Cancer Center Research Institute, Tsukiji 5-1-1, Chuo-ku, Tokyo, 104-0045, Japan. tkiyono@ncc.go.jp.
authors.forename: Tohru
authors.initials: T
authors.lastname: Kiyono
keywords: Cellular conservation; Cultured cells; Endangered animals; Immortalization