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Cytotechnology

, Volume 65, Issue 5, pp 803–809 | Cite as

The transfer of human artificial chromosomes via cryopreserved microcells

  • Narumi Uno
  • Katsuhiro Uno
  • Susi Zatti
  • Kana Ueda
  • Masaharu Hiratsuka
  • Motonobu Katoh
  • Mitsuo OshimuraEmail author
Technical Note

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.

Keywords

Chromosome Human artificial chromosome Microcell-mediated chromosome transfer HAC Cancer Synthetic biology Gene delivery 

Abbreviations

HAC

Human artificial chromosome

PAC

P1 phage-derived artificial chromosome

BAC

Bacterial artificial chromosome

YAC

Yeast artificial chromosome

iPSC

Induced pluripotent stem cell

MSC

Mesenchemal stem cell

PEG

Polyethylene glycol

H

Hemaglutinin

F

Fusion

MMCT

Microcell mediated chromosome transfer

MV-MMCT

Microcell mediated chromosome transfer using Measles virus fusogen

EGFP

Enhanced green fluorescent protein

Bsd

Blasticidin

Notes

Acknowledgments

This study was supported by Regional Innovation Strategy Support Program from The Ministry of Education, Culture, Sports, Science, and Technology of Japan (M. O.), Japan Science and Technology Agency, CREST (M. O.).

Author contributions

N. U. designed and performed most of the experiments. K. U., S. Z., K. U. and H. M. performed the experiments and analyzed the data. M. O. designed the experiments and supervised the entire project. N. U. and M. O. wrote the manuscript.

Conflict of interest

The authors declare no conflicts of interest.

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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Narumi Uno
    • 1
  • Katsuhiro Uno
    • 1
  • Susi Zatti
    • 2
  • Kana Ueda
    • 1
  • Masaharu Hiratsuka
    • 1
  • Motonobu Katoh
    • 1
  • Mitsuo Oshimura
    • 1
    • 3
    Email author
  1. 1.Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medical ScienceTottori UniversityTottoriJapan
  2. 2.Department of Industrial EngineeringUniversity of PadovaPaduaItaly
  3. 3.Chromosome Engineering Research CenterTottori UniversityTottoriJapan

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