Reprogramming of bone marrow derived mesenchymal stromal cells to human induced pluripotent stem cells from pediatric patients with hematological diseases using a commercial mRNA kit

• Published in: Blood cells, molecules, & diseases molecules, & diseases, 2019-05, Vol.76, p.32-39
• Main Authors: Sfougataki, Irene, Grafakos, Ioannis, Varela, Ioanna, Mitrakos, Anastasios, Karagiannidou, Angeliki, Tzannoudaki, Marianna, Poulou, Myrto, Mertzanian, Anny, Roubelakis G., Maria, Stefanaki, Kalliope, Traeger-Synodinos, Joanne, Kanavakis, Emmanuel, Kitra, Vasiliki, Tzetis, Maria, Goussetis, Evgenios
• Format: Article
• Language: English
• Subjects: Disease-specific; Induced pluripotent stem cells; Integration-free; mRNA reprogramming; Pediatric hematological patients
• ISSN: 1079-9796; 1096-0961
• DOI: 10.1016/j.bcmd.2019.01.003

The potential use of patient-specific induced pluripotent stem cells (hiPSCs) in the study and treatment of hematological diseases requires the setup of efficient and safe protocols for hiPSC generation. We aimed to adopt a reprogramming method for large-scale production of integration-free patient-specific hiPSC-lines in our stem cell processing laboratory, which supports a pediatric hematopoietic stem cell transplant unit located at a tertiary care children's hospital. We describe our 5-year experience in generation of hiPSC-lines from human bone marrow-derived mesenchymal stromal cells (BM-MSCs) using synthetic mRNAs encoding reprogramming factors. We generated hiPSC-lines from pediatric patients with β-Thalassemia, Sickle Cell Anemia, Blackfan-Diamond Anemia, Severe Aplastic Anemia, DOCK8 Immunodeficiency and 1 healthy control. After optimization of the reprogramming procedure, average reprogramming efficiency of BM-MSCs was 0.29% (range 0.25–0.4). The complete reprogramming process lasted 14–16 days. Three to five hiPSC-colonies per sample were selected, expanded to 5 culture passages and then frozen. The whole procedure took an average time of 1.8 months (range 1.6–2.2). The hiPSC-lines expressed embryonic stem cell markers and exhibited pluripotency. This mRNA reprogramming method can be applicable in a hematopoietic stem cell culture lab setting and would be useful for the clinical translation of patient-specific hiPSCs.