title: Amniocytes can serve a dual function as a source of iPS cells and feeder layers.
abstract: Clinical barriers to stem-cell therapy include the need for efficient derivation of histocompatible stem cells and the zoonotic risk inherent to human stem-cell xenoculture on mouse feeder cells. We describe a system for efficiently deriving induced pluripotent stem (iPS) cells from human and mouse amniocytes, and for maintaining the pluripotency of these iPS cells on mitotically inactivated feeder layers prepared from the same amniocytes. Both cellular components of this system are thus autologous to a single donor. Moreover, the use of human feeder cells reduces the risk of zoonosis. Generation of iPS cells using retroviral vectors from short- or long-term cultured human and mouse amniocytes using four factors, or two factors in mouse, occurs in 5-7 days with 0.5% efficiency. This efficiency is greater than that reported for mouse and human fibroblasts using similar viral infection approaches, and does not appear to result from selective reprogramming of Oct4(+) or c-Kit(+) amniocyte subpopulations. Derivation of amniocyte-derived iPS (AdiPS) cell colonies, which express pluripotency markers and exhibit appropriate microarray expression and DNA methylation properties, was facilitated by live immunostaining. AdiPS cells also generate embryoid bodies in vitro and teratomas in vivo. Furthermore, mouse and human amniocytes can serve as feeder layers for iPS cells and for mouse and human embryonic stem (ES) cells. Thus, human amniocytes provide an efficient source of autologous iPS cells and, as feeder cells, can also maintain iPS and ES cell pluripotency without the safety concerns associated with xenoculture.
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Raymond M
authors.initials: RM
authors.lastname: Anchan
authors.forename: Philipp
authors.initials: P
authors.lastname: Quaas
authors.forename: Behzad
authors.initials: B
authors.lastname: Gerami-Naini
authors.forename: Hrishikesh
authors.initials: H
authors.lastname: Bartake
authors.forename: Adam
authors.initials: A
authors.lastname: Griffin
authors.forename: Yilan
authors.initials: Y
authors.lastname: Zhou
authors.forename: Daniel
authors.initials: D
authors.lastname: Day
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Eaton
authors.forename: Liji L
authors.initials: LL
authors.lastname: George
authors.forename: Catherine
authors.initials: C
authors.lastname: Naber
authors.forename: Annick
authors.initials: A
authors.lastname: Turbe-Doan
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Park
authors.forename: Mark D
authors.initials: MD
authors.lastname: Hornstein
authors.forename: Richard L
authors.initials: RL
authors.lastname: Maas
mesh_terms: D000650:Amnion / Q000166:cytology*; D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002454:Cell Differentiation; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D005260:Female; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D050810:Octamer Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D050810:Octamer Transcription Factors