Generation of induced pluripotent stem cells and neural cells from a patient with critical illness polyneuropathy
10.3760/cma.j.issn.1671-0282.2018.02.014
- VernacularTitle:危重病多发性神经病患者诱导性多能干细胞系的建立及向神经细胞的诱导分化
- Author:
Huan FU
1
;
Chunhua HU
;
An XIE
;
Yian ZHAN
Author Information
1. 南昌大学第一附属医院麻醉科
- Keywords:
Critical illness polyneuropathy;
Fibroblasts;
Induced pluripotent stem cells;
Neural differentiation;
Disease model
- From:
Chinese Journal of Emergency Medicine
2018;27(2):177-182
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish disease-associated or cell type relevant neuron model generated from patients with Critical Illness Polyneuropathy (CIP) by making CIP patient-derived induced pluripotent stem (iPS) cell lines and neurons to provide a cell-based disease model of CIP.Methods Skin tissue of CIP patient was obtained clinically,and specific skin fibroblasts were isolated and cultured.The iPS cells were derived from CIP patient by introducing 4 transcription factors,namely Oct4,Klf4,Sox2,c-Myc,into patient-specific fibroblast cells by Millipore's Human STEMCCA Constitutive Polycistronic (OKSM) Lentivirus Kit.Colony morphology,alkaline phosphatase (AP) activity,immunofluorescence staining,quantitative reverse transcription polymerase chain reaction (RT-PCR),and differentiation ability were used to identify the pluripotency of these iPS cell lines.In addition,neurons were derived from these iPS cells by inhibiting SMAD pathway.Results The CIP-iPS cells presenting morphological and growth characteristics of human embryonic stem cell (hES) showed the presence of alkaline phosphatase detected by histochemical staining,and the expression of ESC-marker genes.The relative expressions of endogenous pluripotency genes,namely Sox2,REX1,NANOG and OCT4,in iPS cell lines were significantly increased compared with their primary fibroblasts (t values were-9.020,-10.753,-13.295,-12.677,P＜0.01).Subcutaneous injection of iPS cells into NOD-SCID mice resulted in teratomas containing tissues from all the 3 germ layers.Furthermore,cholinergic neurons were successfully induced from CIP-iPS cells.Conclusion The CIP patient-specific iPS cell line and cholinergic neurons were successfully established.Furthermore,the CIP-iPS cell line can be used as models for further elucidating the cellular pathology and developing therapeutic strategies for Critical Illness Polyneuropathy.
