Generation of integration-free iPS cell lines from three sickle cell disease patients from the state of Bahia, Brazil

Martins GLS, Paredes BD, Azevedo CM, Sampaio GLA, Nonaka CKV, Cavalcante BRR, Da Silva KN, Pereira CSE, Soares MBP, Dos Santos RR, Souza BSF
Source: Stem Cell Res
Publication Date: (2018)
Issue: 33: 10-14
Research Area:
Immunotherapy / Hematology
Stem Cells
Cells used in publication:
PBMC, human
Species: human
Tissue Origin: blood
Induced Pluripotent Stem Cell (iPS), human
Species: human
Tissue Origin:
4D-Nucleofector™ X-Unit

Authors used 4D Nucleofector to reprogram erythroblasts into iPS. Episomal vectors from Addgene were nucleofected using P3 Solution and pulse EO-100. 


Sickle cell disease (SCD) is one of the most prevalent and severe monogenetic disorders, affecting several million people around the world. Clinical manifestations
and complications of the disease include sickle cell pain crisis, silent cerebral infarct, stroke, nephropathy and early death. In this study, we generated induced
pluripotent stem cell (iPSC) lines from three homozygous SCD patients from the state of Bahia, Brazil, where SCD is highly prevalent. Peripheral blood mononuclear
cells were collected and erythroblasts were expanded for cell reprogramming with the use of non-integrative episomal vectors. The generated iPSC lines expressed
high levels of pluripotency markers, presented a normal karyotype and were able to differentiate into the three germ layers in embryoid body spontaneous differentiation
assays. Moreover, the expression of the episomal vectors was lost in all iPSC lines after 15 passages. These iPSC lines may help increasing the knowledge
about SCD pathogenesis and can be a useful tool for drug testing and gene editing studies.