title: The transfer of human artificial chromosomes via cryopreserved microcells.
abstract: Microcell-mediated chromosome transfer (MMCT) technology enables a single and intact mammalian chromosome or megabase-sized chromosome fragments to be transferred from donor to recipient cells. The conventional MMCT method is performed immediately after the purification of microcells. The timing of the isolation of microcells and the preparation of recipient cells is very important. Thus, ready-made microcells can improve and simplify the process of MMCT. Here, we established a cryopreservation method to store microcells at -80 °C, and compared these cells with conventionally- (immediately-) prepared cells with respect to the efficiency of MMCT and the stability of a human artificial chromosome (HAC) transferred to human HT1080 cells. The HAC transfer in microcell hybrids was confirmed by FISH analysis. There was no significant difference between the two methods regarding chromosome transfer efficiency and the retention rate of HAC. Thus, cryopreservation of ready-to-use microcells provides an improved and simplified protocol for MMCT.
authors.affiliation: Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medical Science, Tottori University, 86 Nishi-cho, Yonago, Tottori, 683-8503, Japan.
authors.forename: Narumi
authors.initials: N
authors.lastname: Uno
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Uno
authors.forename: Susi
authors.initials: S
authors.lastname: Zatti
authors.forename: Kana
authors.initials: K
authors.lastname: Ueda
authors.forename: Masaharu
authors.initials: M
authors.lastname: Hiratsuka
authors.forename: Motonobu
authors.initials: M
authors.lastname: Katoh
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Oshimura