A validated cellular biobank for β-thalassemia.

Autor:	Cosenza LC; Department of Life Sciences and Biotechnology, Section of Biochemistry and Molecular Biology, University of Ferrara, Via Fossato di Mortara 74, 44121, Ferrara, Italy., Breda L; Department of Hematology-Oncology, Weill Cornell Medical College, New York, NY, USA. bredal@email.chop.edu.; Department of Hematology, Children's Hospital of Philadelphia, 3615 Civic Center Blvd, Abramson Research Center Philadelphia, Philadelphia, PA, 19104, USA. bredal@email.chop.edu., Breveglieri G; Laboratory for the Development of Gene and Pharmacogenomic Therapy of Thalassemia, Biotechnology Centre of Ferrara University, Ferrara, Italy., Zuccato C; Department of Life Sciences and Biotechnology, Section of Biochemistry and Molecular Biology, University of Ferrara, Via Fossato di Mortara 74, 44121, Ferrara, Italy., Finotti A; Department of Life Sciences and Biotechnology, Section of Biochemistry and Molecular Biology, University of Ferrara, Via Fossato di Mortara 74, 44121, Ferrara, Italy., Lampronti I; Department of Life Sciences and Biotechnology, Section of Biochemistry and Molecular Biology, University of Ferrara, Via Fossato di Mortara 74, 44121, Ferrara, Italy., Borgatti M; Department of Life Sciences and Biotechnology, Section of Biochemistry and Molecular Biology, University of Ferrara, Via Fossato di Mortara 74, 44121, Ferrara, Italy., Chiavilli F; Servizio di Immunoematologia e Trasfusione, S.I.T., ULSS 18, Rovigo, Italy., Gamberini MR; Dipartimento di Scienze Mediche-Pediatria, Università di Ferrara, Ferrara, Italy., Satta S; Clinica Pediatrica 2a, Dipartimento di Sanità Pubblica, Medicina Clinica e Molecolare, Università di Cagliari, Ospedale Regionale Microcitemie ASL8, Cagliari, Italy., Manunza L; Clinica Pediatrica 2a, Dipartimento di Sanità Pubblica, Medicina Clinica e Molecolare, Università di Cagliari, Ospedale Regionale Microcitemie ASL8, Cagliari, Italy., De Martis FR; Clinica Pediatrica 2a, Dipartimento di Sanità Pubblica, Medicina Clinica e Molecolare, Università di Cagliari, Ospedale Regionale Microcitemie ASL8, Cagliari, Italy., Moi P; Clinica Pediatrica 2a, Dipartimento di Sanità Pubblica, Medicina Clinica e Molecolare, Università di Cagliari, Ospedale Regionale Microcitemie ASL8, Cagliari, Italy., Rivella S; 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, Philadelphia, PA, 19104, USA., Gambari R; Department of Life Sciences and Biotechnology, Section of Biochemistry and Molecular Biology, University of Ferrara, Via Fossato di Mortara 74, 44121, Ferrara, Italy. gam@unife.it.; Laboratory for the Development of Gene and Pharmacogenomic Therapy of Thalassemia, Biotechnology Centre of Ferrara University, Ferrara, Italy. gam@unife.it., Bianchi N; Department of Life Sciences and Biotechnology, Section of Biochemistry and Molecular Biology, University of Ferrara, Via Fossato di Mortara 74, 44121, Ferrara, Italy.
Jazyk:	angličtina
Zdroj:	Journal of translational medicine [J Transl Med] 2016 Sep 02; Vol. 14, pp. 255. Date of Electronic Publication: 2016 Sep 02.
DOI:	10.1186/s12967-016-1016-4
Abstrakt:	Background: Cellular 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. Methods: Erythroid 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. Results: In 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. Conclusion: The 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.
Databáze:	MEDLINE
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