A validated cellular biobank for β-thalassemia

被引：23

作者：

Cosenza, Lucia Carmela ^{[1
]}

Breda, Laura ^{[2
,3
]}

Breveglieri, Giulia ^{[4
]}

Zuccato, Cristina ^{[1
]}

Finotti, Alessia ^{[1
]}

Lampronti, Ilaria ^{[1
]}

Borgatti, Monica ^{[1
]}

Chiavilli, Francesco ^{[5
]}

Gamberini, Maria Rita ^{[6
]}

Satta, Stefania ^{[7
]}

Manunza, Laura ^{[7
]}

De Martis, Franca Rosa ^{[7
]}

Moi, Paolo ^{[7
]}

Rivella, Stefano ^{[2
,3
]}

Gambari, Roberto ^{[1
,4
]}

Bianchi, Nicoletta ^{[1
]}

机构：

[1] Univ Ferrara, Sect Biochem & Mol Biol, Dept Life Sci & Biotechnol, Via Fossato di Mortara 74, I-44121 Ferrara, Italy

[2] Weill Cornell Med Coll, Dept Hematol Oncol, New York, NY 10065 USA

[3] Childrens Hosp Philadelphia, Dept Hematol, Abramson Res Ctr Philadelphia, 3615 Civ Ctr Blvd, Philadelphia, PA 19104 USA

[4] Univ Ferrara, Ctr Biotechnol, Lab Dev Gene & Pharmacogen Therapy Thalassemia, Ferrara, Italy

[5] ULSS 18, Serv Immunoematol & Trasfus, Rovigo, Italy

[6] Univ Ferrara, Dipartimento Sci Med Pediat, Ferrara, Italy

[7] Univ Cagliari, Osped Reg Microcitemie ASL8, Dipartimento Sanita Pubbl Med Clin & Mol, Clin Pediat 2a, Cagliari, Italy

来源：

JOURNAL OF TRANSLATIONAL MEDICINE | 2016年 / 14卷

关键词：

Thalassemia; Biobanking; HbF induction; Gene therapy; HUMAN ERYTHROID PROGENITORS; CULTURE; HYDROXYUREA; INTEGRATION; DESIGN; CELLS;

D O I：

10.1186/s12967-016-1016-4

中图分类号：

R-3 [医学研究方法]; R3 [基础医学];

学科分类号：

1001 ;

摘要：

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 beta-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 beta-thalassemic, expanded and cryo-preserved. 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 beta-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.

引用

页数：13

共 32 条

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