A validated cellular biobank for β-thalassemia.
A validated cellular biobank for β-thalassemia.
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DOI：
10.1186/s12967-016-1016-4
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发表时间：
2016-09-02
影响因子：
7.4
通讯作者：
Bianchi N
中科院分区：
文献类型：
--
作者：
Cosenza LC;Breda L;Breveglieri G;Zuccato C;Finotti A;Lampronti I;Borgatti M;Chiavilli F;Gamberini MR;Satta S;Manunza L;De Martis FR;Moi P;Rivella S;Gambari R;Bianchi N
细胞生物库是计划在旨在解决各种生物学和生物医学问题的研究中使用相同细胞的协作网络的关键资源。这种方法在β-地中海贫血研究中非常重要，因为患者的招募和标本的收集可能是实验计划中的关键且通常是限制因素。从72名患者中获得红系前体细胞，大多数是β-地中海贫血，扩增并冷冻保存。通过真实的RT-qPCR分析球蛋白基因的表达。通过HPLC研究血红蛋白的产生。在本文中，我们描述了由扩增的红系前体细胞从β-地中海贫血患者构成的Thal-Biobank的生产和验证。在（a）独立采血期间从相同患者中分离细胞，（B）在不同生物库冷冻管中进行冷冻步骤，（c）在不同时间点进行解冻步骤和分析后，验证生物库样本表型的维持。通过将冷冻的生物库细胞运送到不同的实验室，在那里使用相同的标准化程序解冻、培养和分析细胞来确认重现性。根据胎儿血红蛋白产生的基线水平对生物库细胞进行分层，并暴露于胎儿血红蛋白诱导剂。生物库细胞的使用允许对患者的胎儿血红蛋白产生进行分层，并可用于确定对胎儿血红蛋白诱导剂羟基脲和基因治疗方案的反应，结果可重现。本文的在线版本（doi：10.1186/s12967-016-1016-4）包含补充材料，可供授权用户使用。
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. 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. 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. 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. The online version of this article (doi:10.1186/s12967-016-1016-4) contains supplementary material, which is available to authorized users.
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DOI：
10.1073/pnas.1512076112
发表时间：
2015-08-04
影响因子：
11.1
作者：
Mori, Yasuo;Chen, James Y.;Weissman, Irving L.
通讯作者：
Weissman, Irving L.
影响因子：
17.3
作者：
Gaj, Thomas;Gersbach, Charles A.;Barbas, Carlos F., III
通讯作者：
Barbas, Carlos F., III
影响因子：
3.1
作者：
Fibach, Eitan;Bianchi, Nicoletta;Gambari, Roberto
通讯作者：
Gambari, Roberto
影响因子：
3.7
作者：
Dong S;Lin J;Held NL;Clem RJ;Passarelli AL;Franz AW
通讯作者：
Franz AW
影响因子：
3.6
作者：
Fibach, E
通讯作者：
Fibach, E