A validated cellular biobank for β-thalassemia

Journal of Translational Medicine

Research

Paolo Moi¹  Franca Rosa De Martis¹  Laura Manunza¹  Stefania Satta¹  Laura Breda²  Stefano Rivella²  Alessia Finotti³  Monica Borgatti³  Cristina Zuccato³  Nicoletta Bianchi³  Lucia Carmela Cosenza³  Ilaria Lampronti³  Roberto Gambari⁴  Maria Rita Gamberini⁵  Giulia Breveglieri⁶  Francesco Chiavilli⁷ 

[1] Clinica Pediatrica 2a, Dipartimento di Sanità Pubblica, Medicina Clinica e Molecolare, Università di Cagliari, Ospedale Regionale Microcitemie ASL8, Cagliari, Italy;Department of Hematology-Oncology, Weill Cornell Medical College, New York, NY, USA;Department of Hematology, Children’s Hospital of Philadelphia, 3615 Civic Center Blvd, Abramson Research Center Philadelphia, 19104, Philadelphia, PA, USA;Department of Life Sciences and Biotechnology, Section of Biochemistry and Molecular Biology, University of Ferrara, Via Fossato di Mortara 74, 44121, Ferrara, Italy;Department of Life Sciences and Biotechnology, Section of Biochemistry and Molecular Biology, University of Ferrara, Via Fossato di Mortara 74, 44121, Ferrara, Italy;Laboratory for the Development of Gene and Pharmacogenomic Therapy of Thalassemia, Biotechnology Centre of Ferrara University, Ferrara, Italy;Dipartimento di Scienze Mediche-Pediatria, Università di Ferrara, Ferrara, Italy;Laboratory for the Development of Gene and Pharmacogenomic Therapy of Thalassemia, Biotechnology Centre of Ferrara University, Ferrara, Italy;Servizio di Immunoematologia e Trasfusione, S.I.T., ULSS 18, Rovigo, Italy;

关键词: Thalassemia;    Biobanking;    HbF induction;    Gene therapy;   

DOI  :  10.1186/s12967-016-1016-4

 received in 2016-05-20, accepted in 2016-08-18,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundCellular biobanking is a key resource for collaborative networks planning to use same cells in studies aimed at solving a variety of biological and biomedical issues. This approach is of great importance in studies on β-thalassemia, since the recruitment of patients and collection of specimens can represent a crucial and often limiting factor in the experimental planning.MethodsErythroid precursor cells were obtained from 72 patients, mostly β-thalassemic, expanded and cryopreserved. Expression of globin genes was analyzed by real time RT-qPCR. Hemoglobin production was studied by HPLC.ResultsIn this paper we describe the production and validation of a Thal-Biobank constituted by expanded erythroid precursor cells from β-thalassemia patients. The biobanked samples were validated for maintenance of their phenotype after (a) cell isolation from same patients during independent phlebotomies, (b) freezing step in different biobanked cryovials, (c) thawing step and analysis at different time points. Reproducibility was confirmed by shipping the frozen biobanked cells to different laboratories, where the cells were thawed, cultured and analyzed using the same standardized procedures. The biobanked cells were stratified on the basis of their baseline level of fetal hemoglobin production and exposed to fetal hemoglobin inducers.ConclusionThe use of biobanked cells allows stratification of the patients with respect to fetal hemoglobin production and can be used for determining the response to the fetal hemoglobin inducer hydroxyurea and to gene therapy protocols with reproducible results.

【 授权许可】

CC BY   
© The Author(s) 2016