Establishment of human iPSC line from patient of Indian ethnicity carrying homozygous CD8/9 (+G) beta thalassemia mutation

Shruti Tembe; Sophia Fernandes; Nikhat Khan; Sameer Melinkeri; Vaijayanti Kale; Lalita Limaye

doi:10.1016/j.scr.2020.101772

Establishment of human iPSC line from patient of Indian ethnicity carrying homozygous CD8/9 (+G) beta thalassemia mutation

Stem Cell Res. 2020 Apr:44:101772. doi: 10.1016/j.scr.2020.101772. Epub 2020 Mar 20.

Authors

Shruti Tembe¹, Sophia Fernandes¹, Nikhat Khan¹, Sameer Melinkeri², Vaijayanti Kale¹, Lalita Limaye³

Affiliations

¹ Stem Cell Laboratory, National Centre for Cell Science, NCCS Complex, University of Pune Campus, Ganeshkhind, Pune 411007, India.
² Blood and Marrow Transplant Unit, Deenanath Mangeshkar Hospital, Erandwane, Pune 411004, India.
³ Stem Cell Laboratory, National Centre for Cell Science, NCCS Complex, University of Pune Campus, Ganeshkhind, Pune 411007, India. Electronic address: [email protected].

PMID: 32278313
DOI: 10.1016/j.scr.2020.101772

Abstract

This study shows generation of iPSCs from peripheral blood mononuclear cells (PBMNCs) of a male patient having homozygous CD 8/9 (+G) beta thalassemia (major) mutation. Cells were nucleofected with episomal vectors containing Oct4, Sox2, L-Myc, Lin28, Klf4 and p53DD (dominant negative p53 mutation). Cell line exhibited presence of pluripotency markers by immunofluorescence, flow-cytometry and PCR. The plasmids were lost from cells by subsequent passages, observed by PCR. Karyotype analysis demonstrated a stable genome. The cells had capability to differentiate into three-germ lineages in vitro. This iPSC line can be used as a tool for drug design and gene therapy studies.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CD8-Positive T-Lymphocytes
Cell Differentiation
Ethnicity
Humans
Induced Pluripotent Stem Cells*
Kruppel-Like Factor 4
Leukocytes, Mononuclear
Male
Mutation
beta-Thalassemia* / genetics