AI Article Synopsis

  • Synthetic modified mRNA molecules have been successfully used to reprogram human fibroblasts into induced pluripotent stem cells (iPSCs) and have now been applied to bone marrow-derived mesenchymal stromal cells (BM-MSCs) from a β-thalassemia patient to create transgene-free β-thal-iPSCs.
  • Transfection of BM-MSCs with specific reprogramming factors resulted in the formation of iPSC colonies, which maintained genetic stability and displayed characteristics similar to human embryonic stem cells (ESCs) after several passages.
  • The generated β-thal-iPSCs show potential for applications in disease modeling and gene therapy, demonstrating that mRNA-based reprogramming is effective beyond just fibroblasts

Article Abstract

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 10(4) 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.

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http://dx.doi.org/10.1089/cell.2014.0050DOI Listing

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