Derivation of induced pluripotent stem cells by retroviral gene transduction in mammalian species.
Pluripotent stem cells can provide us with an enormous cell source for in vitro model systems for development. In 2006, new methodology was designed to generate pluripotent stem cells directly from somatic cells, and these cells were named induced pluripotent stem cells (iPSCs). This method consists of technically simple procedures: donor cell preparation, gene transduction, and isolation of embryonic stem cell-like colonies. The iPSC technology enables cell biologists not only to obtain pluripotent stem cells easily but also to study the reprogramming events themselves. Here, we describe the protocols to generate iPSCs from somatic origins by using conventional viral vectors. Specifically, we state the usage of three mammalian species: mouse, common marmoset, and human. As mouse iPSC donors, fibroblasts are easily prepared, while mesenchymal stem cells are expected to give rise to highly reprogrammed iPSCs efficiently. Common marmoset (Callithrix jacchus), a nonhuman primate, represents an alternative model to the usual laboratory animals. Finally, patient-specific human iPSCs give us an opportunity to examine the pathology and mechanisms of dysregulated genomic imprinting. The iPSC technology will serve as a valuable method for studying genomic imprinting, and conversely, the insights from these studies will offer valuable criteria to assess the potential of iPSCs.
Department of Physiology, School of Medicine, Keio University, Tokyo, Japan.
Masanori
M
Imamura
Hironobu
H
Okuno
Ikuo
I
Tomioka
Yoshimi
Y
Kawamura
Zachary Yu-Ching
ZY
Lin
Ryusuke
R
Nakajima
Wado
W
Akamatsu
Hirotaka James
HJ
Okano
Yumi
Y
Matsuzaki
Erika
E
Sasaki
Hideyuki
H
Okano
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D002144:Callithrix; D018929:Cell Culture Techniques; D002469:Cell Separation; D015925:Cryopreservation; D004622:Embryo, Mammalian / Q000166:cytology; D005260:Female; D005333:Fetus / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology; D018392:Genomic Imprinting; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D016086:Lentivirus / Q000235:genetics / Q000502:physiology; D008099:Liver / Q000166:cytology; D059630:Mesenchymal Stem Cells / Q000166:cytology; D051379:Mice; D011247:Pregnancy; D012190:Retroviridae / Q000235:genetics* / Q000502:physiology; D013623:Tail / Q000166:cytology; D014161:Transduction, Genetic / Q000379:methods*