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.
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 immunodeficientmice. 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: Andrea R Argouarch; Nina Schultz; Andrew C Yang; Yeongjun Jang; Kristle Garcia; Celica G Cosme; Christian I Corrales; Alissa L Nana; Anna M Karydas; Salvatore Spina; Lea T Grinberg; Bruce Miller; Tony Wyss-Coray; Alexej Abyzov; Hani Goodarzi; William W Seeley; Aimee W Kao Journal: Stem Cell Rev Rep Date: 2022-07-09 Impact factor: 6.692
Authors: Missy T Pham; Kari M Pollock; Melanie D Rose; Whitney A Cary; Heather R Stewart; Ping Zhou; Jan A Nolta; Ben Waldau Journal: Neuroreport Date: 2018-05-02 Impact factor: 1.837
Authors: Kaitlin C Clark; Fernando A Fierro; Emily Mills Ko; Naomi J Walker; Boaz Arzi; Clifford G Tepper; Heather Dahlenburg; Andrew Cicchetto; Amir Kol; Lyndsey Marsh; William J Murphy; Nasim Fazel; Dori L Borjesson Journal: Stem Cell Res Ther Date: 2017-03-20 Impact factor: 6.832
Authors: María Lacalle-Aurioles; Camille Cassel de Camps; Cornelia E Zorca; Lenore K Beitel; Thomas M Durcan Journal: Front Cell Neurosci Date: 2020-11-12 Impact factor: 5.505