An integration-free iPSC line (SDQLCHi017-A) derived from a patient with nemaline myopathy-2 disease carrying compound heterozygote mutations in NEB gene

Authors:
Yanyan Ma, Haiyan Zhang, Xiaomei Li, Xiaomeng Yang, Yue Li, Jingyun Guan, Yuqiang Lv, Zhongtao Gai, Yi Liu
In:
Source: Stem Cell Res
Publication Date: (2020)
Issue: 43: 101729
Research Area:
Stem Cells
Cells used in publication:
PBMC, human
Species: human
Tissue Origin: blood
Platform:
4D-Nucleofector™ X-Unit
Experiment

PBMCs were isolated from patient’s peripheral blood and cultured for some time to enrich erythroblasts. Solution P3 and EO-100 and non-integrating vectors were used for reprogramming. After reprogramming, cells were cultivated on feeder (MEF). First colonies isolated after 2 weeks.

Abstract

Nemaline myopathy-2 (NEM2) is an autosomal recessive skeletal muscle disorder caused by mutations in the nebulin (NEB) gene. We report the generation and characterization of a human induced pluripotent stem cell (iPSC) line SDQLCHi017-A, derived from a 1-month-old patient with NEM2 carrying compound heterozygote mutations (c.6915+1G>T, c.14910+3G>C) in NEB gene. The peripheral blood mononuclear cells (PBMCs) were reprogrammed with non-integrating episomal vectors coding OCT4, SOX2, KLF4, BCL-XL and MYC. The established iPSC line contained the same mutations found in the patient, showed a normal karyotype, could differentiate into cells of three germ layers in vitro and expressed pluripotency markers.