title: Efficient Generation of Induced Pluripotent Stem and Neural Progenitor Cells From Acutely Harvested Dura Mater Obtained During Ventriculoperitoneal Shunt Surgery.
abstract: BACKGROUND
  The dura mater can be easily biopsied during most cranial neurosurgical operations. We describe a protocol that allows for robust generation of induced pluripotent stem cells (iPSCs) and neural progenitors from acutely harvested dura mater.
  
  
  OBJECTIVE
  To generate iPSCs and neural progenitor cells from dura mater obtained during ventriculoperitoneal shunt surgery.
  
  
  METHODS
  Dura was obtained during ventriculoperitoneal shunt surgery for normal pressure hydrocephalus from a 60-year-old patient with severe cognitive impairment. Fibroblasts were isolated from the dural matrix and transduced with nonintegrating Sendai virus for iPSC induction. A subset of successfully generated iPSC clones underwent immunocytochemical analysis, teratoma assay, karyotyping, and targeted neural differentiation.
  
  
  RESULTS
  Eleven iPSC clones were obtained from the transduction of an estimated 600,000 dural fibroblasts after 3 passages. Three clones underwent immunocytochemical analysis and were shown to express the transcription factors OCT-4, SOX2, and the embryonic cell markers SSEA-4, TRA-1-60, and Nanog. Two clones were tested for pluripotency and formed teratomas at the injection site in immunodeficient mice. Three clones underwent chromosomal analysis and were found to have a normal metaphase spread and karyotype. One clone underwent targeted neural differentiation and formed neural rosettes as well as TuJ1/SOX1-positive neural progenitor cells.
  
  
  CONCLUSIONS
  IPSCs and neural progenitor cells can be efficiently derived from the dura of patients who need to undergo cranial neurosurgical operations. IPSCs were obtained with a nonintegrating virus and exhibited a normal karyotype, making them candidates for future autotransplantation after targeted differentiation to treat functional deficits.
authors.affiliation: Institute for Regenerative Cures (IRC), Sacramento, California, USA; UC Davis Stem Cell Program, Sacramento, California, USA.
authors.forename: Whitney Ann
authors.initials: WA
authors.lastname: Cary
authors.affiliation: Institute for Regenerative Cures (IRC), Sacramento, California, USA; UC Davis Stem Cell Program, Sacramento, California, USA.
authors.forename: Courtney Namiko
authors.initials: CN
authors.lastname: Hori
authors.affiliation: Institute for Regenerative Cures (IRC), Sacramento, California, USA; UC Davis Stem Cell Program, Sacramento, California, USA.
authors.forename: Missy Trananh
authors.initials: MT
authors.lastname: Pham
authors.affiliation: Institute for Regenerative Cures (IRC), Sacramento, California, USA; UC Davis Stem Cell Program, Sacramento, California, USA.
authors.forename: Catherine Ann
authors.initials: CA
authors.lastname: Nacey
authors.affiliation: Institute for Regenerative Cures (IRC), Sacramento, California, USA; UC Davis Stem Cell Program, Sacramento, California, USA.
authors.forename: Jeannine Logan
authors.initials: JL
authors.lastname: McGee
authors.affiliation: Institute for Regenerative Cures (IRC), Sacramento, California, USA; UC Davis Stem Cell Program, Sacramento, California, USA.
authors.forename: Mattan
authors.initials: M
authors.lastname: Hamou
authors.affiliation: Department of Neurosurgery, UC Davis Medical Center, Sacramento, California, USA.
authors.forename: Robert F
authors.initials: RF
authors.lastname: Berman
authors.affiliation: Institute for Regenerative Cures (IRC), Sacramento, California, USA; UC Davis Stem Cell Program, Sacramento, California, USA.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Bauer
authors.affiliation: Institute for Regenerative Cures (IRC), Sacramento, California, USA; UC Davis Stem Cell Program, Sacramento, California, USA.
authors.forename: Jan A
authors.initials: JA
authors.lastname: Nolta
authors.affiliation: Department of Neurosurgery, UC Davis Medical Center, Sacramento, California, USA; UC Davis Stem Cell Program, Sacramento, California, USA. Electronic address: ben.waldau@ucdmc.ucdavis.edu.
authors.forename: Ben
authors.initials: B
authors.lastname: Waldau
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D065150:Cellular Reprogramming; D002999:Clone Cells; D003072:Cognition Disorders / Q000209:etiology; D004388:Dura Mater / Q000166:cytology* / Q000601:surgery; D005347:Fibroblasts; D006801:Humans; D006850:Hydrocephalus, Normal Pressure / Q000601:surgery; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism; D007621:Karyotyping; D008297:Male; D051379:Mice; D008875:Middle Aged; D058953:Neural Stem Cells / Q000378:metabolism; D029082:Sendai virus; D013724:Teratoma / Q000473:pathology; D014157:Transcription Factors / Q000378:metabolism; D017287:Ventriculoperitoneal Shunt
chemical_list: D014157:Transcription Factors
keywords: Dura; Fibroblast; Induced pluripotent stem cell; Neural progenitor cell; Regenerative medicine; Shunt surgery