TY - JOUR
TI - Generation of human β-thalassemia induced pluripotent cell lines by reprogramming of bone marrow-derived mesenchymal stromal cells using modified mRNA
AU - Varela, I.
AU - Karagiannidou, A.
AU - Oikonomakis, V.
AU - Tzetis, M.
AU - Tzanoudaki, M.
AU - Siapati, E.-K.
AU - Vassilopoulos, G.
AU - Graphakos, S.
AU - Kanavakis, E.
AU - Goussetis, E.
JO - Cellular Reprogramming
PY - 2014
VL - 16
TODO - 6
SP - 447-455
PB - MARY ANN LIEBERT INC PUBL
SN - null
TODO - 10.1089/cell.2014.0050
TODO - CD34 antigen;  kruppel like factor 4;  Lin28 protein;  messenger RNA;  Myc protein;  octamer transcription factor 4;  transcription factor;  transcription factor Sox2;  unclassified drug;  messenger RNA;  transcription factor, animal experiment;  animal model;  animal tissue;  Article;  beta thalassemia;  bone marrow derived mesenchymal stem cell;  cell differentiation;  cell lineage;  colony formation;  comparative genomic hybridization;  controlled study;  embryoid body;  embryonic stem cell;  gene expression profiling;  genetic stability;  genetic transfection;  human;  human cell;  in vitro study;  in vivo study;  mouse;  nonhuman;  pluripotent stem cell;  teratoma;  transgene;  cell line;  cytology;  fibroblast;  genetics;  mesenchymal stroma cell;  nuclear reprogramming;  pluripotent stem cell, beta-Thalassemia;  Cell Differentiation;  Cell Line;  Cellular Reprogramming;  Comparative Genomic Hybridization;  Fibroblasts;  Humans;  Induced Pluripotent Stem Cells;  Mesenchymal Stromal Cells;  RNA, Messenger;  Transcription Factors
TODO - Synthetic modified mRNA molecules encoding pluripotency transcription factors have been used successfully in reprogramming human fibroblasts to induced pluripotent stem cells (iPSCs). We have applied this method on bone marrow-derived mesenchymal stromal cells (BM-MSCs) obtained from a patient with β-thalassemia (β-thal) with the aim to generate trangene-free β-thal-iPSCs. Transfection of 104 BM-MSCs by lipofection with mRNA encoding the reprogramming factors Oct4, Klf4, Sox2, cMyc, and Lin28 resulted in formation of five iPSC colonies, from which three were picked up and expanded in β-thal-iPSC lines. After 10 serial passages in vitro, β-thal-iPSCs maintain genetic stability as shown by array comparative genomic hybridization (aCGH) and are capable of forming embryoid bodies in vitro and teratomas in vivo. Their gene expression profile compared to human embryonic stem cells (ESCs) and BM-MSCs seems to be similar to that of ESCs, whereas it differs from the profile of the parental BM-MSCs. Differentiation cultures toward a hematopoietic lineage showed the generation of CD34+ progenitors up to 10%, but with a decreased hematopoietic colony-forming capability. In conclusion, we report herein the generation of transgene-free β-thal-iPSCs that could be widely used for disease modeling and gene therapy applications. Moreover, it was demonstrated that the mRNA-based reprogramming method, used mainly in fibroblasts, is also suitable for reprogramming of human BM-MSCs. © Mary Ann Liebert, Inc. 2014.
ER -