Springer Protocols: Abstract: Nuclear Transfer to Study the Nuclear Reprogramming of Human Stem Cells

Nuclear Transfer to Study the Nuclear Reprogramming of Human Stem Cells

By: Shigeo Saito³, Ken Sawai⁴, Yoshinobu Murayama⁵, Keiichi Fukuda⁶, Kazunari Yokoyama⁷

Abstract

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Research of stem cells will enable us to understand the development and function of tissues and organs in mammals. The ability to induce regeneration of new tissues from embryonic stem (ES) cells derived from cloned blastocysts via nuclear transfer can be expected in the not-too-distant future. The fact that there is no way except nuclear cloning for the return of differentiated cells to undifferentiated cells remains an interesting problem to be solved. We describe protocols for the production of cloned calves from bovine ES cells to study nuclear reprogramming ability of stem cells. The frequency of term pregnancies for blastocysts from ES cells is higher than those of early pregnancies and maintained pregnancies after nuclear transfer with bovine somatic cells. We also describe protocols for gene introduction into bovine ES cells in vitro, particularly the human leukocyte antigens (HLA). Bovine ES cells provide a powerful tool for the generation of transgenic clonal offspring. This technique, when perfected for humans, may be critical for neural stem cell transplantation.

Affiliation(s): (3) Saito Laboratory of Cell Technology, Kataoka, Yaita, Tochigi, Japan
(4) Hokkaido Animal Research Center, Shintoku, Hokkaido, Japan
(5) College of Engineering, Nihon University, Koriyama, Fukushima, Japan
(6) Keio University School of Medicine, Shinjuku, Tokyo, Japan
(7) Gene Engineering Division, Department of Biological Systems, BioResource Center, RIKEN, Tsukuba, Ibaraki, Japan

Book Title: Neural Stem Cells: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 438 | Pub. Date: Feb-01-2008 | Page Range: 151-169 | DOI: 10.1007/978-1-59745-133-8_13

Subject: Cell Biology

Key Words: Nuclear transfer - bovine - ES cell - reprogramming - cloning - stem cells - LIF - HLA - EGF - donor - recipient - transfection - EGFP - pluripotency - differentiation - fusion - microsatellite - PCR - Oct-4 - STAT-3

References

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