title: Cell cloning-on-the-spot by using an attachable silicone cylinder.
abstract: Cell cloning is a laboratory routine to isolate and keep particular properties of cultured cells. Transfected or other genetically modified cells can be selected by the traditional microbiological cloning. In addition, common laboratory cell lines are prone to genotypic drift during their continual culture, so that supplementary cloning steps are often required to maintain correct lineage phenotypes. Here, we designed a silicone-made attachable cloning cylinder, which facilitated an easy and bona fide cloning of interested cells. This silicone cylinder was easy to make, showed competent stickiness to laboratory plastics including culture dishes, and hence enabled secure isolation and culture for days of selected single cells, especially, on the spots of preceding cell-plating dishes under microscopic examination of visible cellular phenotypes. We tested the silicone cylinder in the monoclonal subcloning from a heterogeneous population of a breast cancer cell line, MDA-MB-231, and readily established independent MDA-MB-231 subclones showing different sublineage phenotypes.
authors.affiliation: Department of Biochemistry, College of Medicine, Chungnam National University, Munhwa-ro 266, Jung-gu, Daejeon, 35015, South Korea; Department of Nature Inspired Nano Convergences System, Korea Institute of Machinery and Materials (KIMM), Gajeongbuk-ro 156, Yuseong-gu, 34103, South Korea. Electronic address: garasaki47@nate.com.
authors.forename: Hong Bum
authors.initials: HB
authors.lastname: Park
authors.affiliation: Department of Biochemistry, College of Medicine, Chungnam National University, Munhwa-ro 266, Jung-gu, Daejeon, 35015, South Korea; Research Institute for Medical Sciences, Chungnam National University, Munhwa-ro 266, Jung-gu, Daejeon, 35015, South Korea. Electronic address: sonwonseok@gmail.com.
authors.forename: Wonseok
authors.initials: W
authors.lastname: Son
authors.affiliation: Department of Sculpture, Art School, Chungnam National University, Daehak-ro 99, Yuseong-gu, Daejeon, 34134, South Korea. Electronic address: dong8art@gmail.com.
authors.forename: Dong Han
authors.initials: DH
authors.lastname: Chae
authors.affiliation: Department of Biochemistry, College of Medicine, Chungnam National University, Munhwa-ro 266, Jung-gu, Daejeon, 35015, South Korea. Electronic address: egs0522@naver.com.
authors.forename: Jisu
authors.initials: J
authors.lastname: Lee
authors.affiliation: Department of Biochemistry, College of Medicine, Chungnam National University, Munhwa-ro 266, Jung-gu, Daejeon, 35015, South Korea; Research Institute for Medical Sciences, Chungnam National University, Munhwa-ro 266, Jung-gu, Daejeon, 35015, South Korea. Electronic address: ilwoung@cnu.ac.kr.
authors.forename: Il-Woung
authors.initials: IW
authors.lastname: Kim
authors.affiliation: Department of Biochemistry, College of Medicine, Chungnam National University, Munhwa-ro 266, Jung-gu, Daejeon, 35015, South Korea; Research Institute for Medical Sciences, Chungnam National University, Munhwa-ro 266, Jung-gu, Daejeon, 35015, South Korea. Electronic address: woomizzang@gmail.com.
authors.forename: Woomi
authors.initials: W
authors.lastname: Yang
authors.affiliation: Department of Internal Medicine, College of Medicine, Chungnam National University, Munhwa-ro 266, Jung-gu, Daejeon, 35015, South Korea. Electronic address: jksung69@cnuh.co.kr.
authors.forename: Jae Kyu
authors.initials: JK
authors.lastname: Sung
authors.affiliation: Department of Biochemistry, College of Medicine, Chungnam National University, Munhwa-ro 266, Jung-gu, Daejeon, 35015, South Korea. Electronic address: kyulim@cnu.ac.kr.
authors.forename: Kyu
authors.initials: K
authors.lastname: Lim
authors.affiliation: Department of Nature Inspired Nano Convergences System, Korea Institute of Machinery and Materials (KIMM), Gajeongbuk-ro 156, Yuseong-gu, 34103, South Korea. Electronic address: meek@kimm.re.kr.
authors.forename: Jun Hee
authors.initials: JH
authors.lastname: Lee
authors.affiliation: Department of Pathology, Cancer Research Institute, College of Medicine, Chungnam National University, Munhwa-ro 266, Jung-gu, Daejeon, 35015, South Korea. Electronic address: phone330@cnu.ac.kr.
authors.forename: Kyung-Hee
authors.initials: KH
authors.lastname: Kim
authors.affiliation: Department of Biochemistry, College of Medicine, Chungnam National University, Munhwa-ro 266, Jung-gu, Daejeon, 35015, South Korea; Research Institute for Medical Sciences, Chungnam National University, Munhwa-ro 266, Jung-gu, Daejeon, 35015, South Korea. Electronic address: Jipark@cnu.ac.kr.
authors.forename: Jong-Il
authors.initials: JI
authors.lastname: Park
mesh_terms: D061249:Batch Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002999:Clone Cells; D019976:Cloning, Organism / Q000295:instrumentation* / Q000379:methods*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D012828:Silicones / Q000737:chemistry*
chemical_list: D012828:Silicones
keywords: Cell cloning; MDA-MB-231 breast cancer cell line; Silicone-made cloning cylinder; Single cell cloning