Τίτλος:
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
Γλώσσες Τεκμηρίου:
Αγγλικά
Περίληψη:
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. © 2019 Elsevier Inc.
Συγγραφείς:
Sfougataki, I.
Grafakos, I.
Varela, I.
Mitrakos, A.
Karagiannidou, A.
Tzannoudaki, M.
Poulou, M.
Mertzanian, A.
Roubelakis G., M.
Stefanaki, K.
Traeger-Synodinos, J.
Kanavakis, E.
Kitra, V.
Tzetis, M.
Goussetis, E.
Περιοδικό:
BLOOD CELLS MOLECULES AND DISEASES
Εκδότης:
Academic Press Inc.
Λέξεις-κλειδιά:
biological marker; messenger RNA; messenger RNA, aplastic anemia; Article; beta thalassemia; Blackfan Diamond anemia; bone marrow derived mesenchymal stem cell; cell reprogramming technique; child; controlled study; hematologic disease; human; human cell; immune deficiency; induced pluripotent stem cell; priority journal; process optimization; sickle cell anemia; tertiary care center; cytology; drug effect; hematologic disease; hematopoietic stem cell transplantation; induced pluripotent stem cell; mesenchymal stem cell; nuclear reprogramming; personalized medicine; procedures; synthesis; time factor, Cellular Reprogramming; Child; Hematologic Diseases; Hematopoietic Stem Cell Transplantation; Humans; Induced Pluripotent Stem Cells; Mesenchymal Stem Cells; Methods; Precision Medicine; RNA, Messenger; Time Factors
DOI:
10.1016/j.bcmd.2019.01.003
