Reprogramming of patient derived cells is becoming a more and more common methodology for disease studies, especially peripheral blood mononuclear cells (PBMCs). PBMCs are easy to collect and well-established protocols for reprogramming into iPSCs by using Lonza’s NucleofectorTM Devices exist.
Three recent examples for reprogramming of patient derived PBMC into iPSCs are presented here. Cell line CSUASOi004-A was derived from a 15-year-old female patient suffering from a PRPF6 mutation causing Retinitis Pigmentosa (Zhou et al., 2020). Cell line SDQLCHi017-A was derived from a 1-month-old patient with mutations in the NEB gene (Ma et al., 2020), and cell line SDQLCHi007-A was derived from a 4-year-old male patient diagnosed with Duchenne muscular dystrophy caused by mutations in the dystrophin gene (Guan et al., 2020). All these iPSCs were generated by delivering episomal vectors to the patient derived cells. All three generated cell lines still carry the disease mutations and show an iPSC-like morphology, expression of pluripotency markers, and a normal karyotype. Differentiation potential into all three germ layers was confirmed for all lines by qRT-PCR.
These results show that reprogramming of human somatic cells into iPSC by using NucleofectorTM Devices is an easy and helpful method to gain insights into disease mechanisms by using cells collected from patients suffering from these diseases.
Written by SST
References
- Jingyun Guan, Xinnong Liu, Haiyan Zhang, Xiaomeng Yang, Yanyan Ma, Yue Li, Zhongtao Gai, Yi Liu. Reprogramming of human Peripheral Blood Mononuclear Cell (PBMC) from a Chinese patient suffering Duchenne muscular dystrophy to iPSC line (SDQLCHi007-A) carrying deletion of 49-50 exons in the DMD gene. Stem Cell Res. 2020 Jan;42:101666.
- Yanyan Ma, Haiyan Zhang, Xiaomei Li, Xiaomeng Yang, Yue Li, Jingyun Guan, Yuqiang Lv, Zhongtao Gai, Yi Liu. An integration-free iPSC line (SDQLCHi017-A) derived from a patient with nemaline myopathy-2 disease carrying compound heterozygote mutations in NEB gene. Stem Cell Res. 2020 Mar;43:101729.
- Yalan Zhou 1, Yutong Jing 2, Shengru Mao 3, Jian Liu 4, Zekai Cui 5, Yini Wang 6, Juan Chen 7, Hon Fai Chan 8, Shibo Tang 9, Jiansu Chen. Establishment of induced pluripotent stem cell line CSUASOi004-A by reprogramming peripheral blood mononuclear cells of a PRPF6-related dominant retinitis pigmentosa patient. Stem Cell Res. 2020 May;45:101793.
