Efficient Generation of Induced Pluripotent Stem and Neural Progenitor Cells From Acutely Harvested Dura Mater Obtained During Ventriculoperitoneal Shunt Surgery.
Efficient Generation of Induced Pluripotent Stem and Neural Progenitor Cells From Acutely Harvested Dura Mater Obtained During Ventriculoperitoneal Shunt Surgery.
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DOI：
10.1016/j.wneu.2015.05.076
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发表时间：
2015-11
影响因子：
2
通讯作者：
Waldau B
中科院分区：
文献类型：
--
作者：
Cary WA;Hori CN;Pham MT;Nacey CA;McGee JL;Hamou M;Berman RF;Bauer G;Nolta JA;Waldau B
在大多数颅神经外科手术中，可以轻松对硬脑膜进行活检。我们描述了一种协议，允许从急性收获的硬脑膜中大量生成诱导多能干细胞（iPSC）和神经祖细胞。从脑室腹腔分流手术期间获得的硬脑膜中生成 iPSC 和神经祖细胞。硬脑膜是在一名患有严重认知障碍的 60 岁患者的正常压力脑积水的脑室腹腔分流手术中获得的。从硬脑膜基质中分离成纤维细胞，并用非整合仙台病毒转导以诱导 iPSC。对成功生成的 iPSC 克隆的子集进行了免疫细胞化学分析、畸胎瘤测定、核型分析和靶向神经分化。经过 3 次传代后，从估计 600,000 个硬脑膜成纤维细胞的转导中获得了 11 个 iPSC 克隆。三个克隆进行了免疫细胞化学分析，结果显示其表达转录因子 OCT-4、SOX2 以及胚胎细胞标记物 SSEA-4、TRA-1-60 和 Nanog。对两个克隆进行多能性测试，并在免疫缺陷小鼠的注射部位形成畸胎瘤。三个克隆进行了染色体分析，发现具有正常的中期扩散和核型。一个克隆进行了靶向神经分化，并形成了神经玫瑰花结以及 TuJ1/SOX1 阳性神经祖细胞。 IPSCs和神经祖细胞可以有效地从需要接受颅神经外科手术的患者的硬脑膜中提取。 IPSCs是用非整合病毒获得的，并表现出正常的核型，使其成为未来自体移植的候选者，经过定向分化以治疗功能缺陷。
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. To generate iPSCs and neural progenitor cells from dura mater obtained during ventriculoperitoneal shunt surgery. 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. 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. 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.
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影响因子：
3.7
作者：
Suhr ST;Chang EA;Rodriguez RM;Wang K;Ross PJ;Beyhan Z;Murthy S;Cibelli JB
通讯作者：
Cibelli JB
影响因子：
5.9
作者：
Doi, Daisuke;Samata, Bumpei;Katsukawa, Mitsuko;Kikuchi, Tetsuhiro;Morizane, Asuka;Ono, Yuichi;Sekiguchi, Kiyotoshi;Nakagawa, Masato;Parmar, Malin;Takahashi, Jun
通讯作者：
Takahashi, Jun
影响因子：
7.1
作者：
Sproul, Andrew A.;Vensand, Lauren B.;Noggle, Scott A.
通讯作者：
Noggle, Scott A.
影响因子：
4.2
作者：
Nudi, Evan T.;Jacqmain, Justin;Smith, Jeffrey S.
通讯作者：
Smith, Jeffrey S.
影响因子：
64.5
作者：
Takahashi, Kazutoshi;Yamanaka, Shinya
通讯作者：
Yamanaka, Shinya