Roda Barbara, Reschiglian Pierluigi, Zattoni Andrea, Tazzari Pier Luigi, Buzzi Marina, Ricci Francesca, Bontadini Andrea
Department of Chemistry "G. Ciamician", University of Bologna, Via Selmi 2, 40126, Bologna, Italy.
Anal Bioanal Chem. 2008 Sep;392(1-2):137-45. doi: 10.1007/s00216-008-2271-x. Epub 2008 Jul 24.
Interest in biological studies on various cell types for many biomedical applications, from research to patient treatments, is constantly increasing. The ability to discriminate (sort) and/or quantify distinct subpopulations of cells has become increasingly important. For instance, not only detection but also the highest depletion of neoplastic cells from normal cells is an important requisite in the autologous transplantation of lymphocytes for blood cancer treatments. In this work, gravitational field-flow fractionation (GrFFF) is shown to be effective for sorting a heterogeneous mixture of human, living lymphocytes constituted of neoplastic B cells from a Burkitt lymphoma cell line and healthy T and B lymphocytes from blood samples. GrFFF does not require the use of fluorescent immunotags for sorting cells, and the sorted cells can be collected for their further characterization. Flow cytometry was used to assess the viability of the cells collected, and to evaluate the cell fractionation achieved. A low amount of neoplastic B lymphocytes (less than 2%) was found in a specific fraction obtained by GrFFF. The high depletion from neoplastic cells (more than 98%) was confirmed by a clonogenicity test.
从研究到患者治疗，在许多生物医学应用中，对各种细胞类型进行生物学研究的兴趣正在不断增加。区分（分选）和/或量化不同细胞亚群的能力变得越来越重要。例如，在用于血癌治疗的淋巴细胞自体移植中，不仅检测而且从正常细胞中最大程度地去除肿瘤细胞都是一项重要要求。在这项工作中，重力场流分离法（GrFFF）被证明可有效分选由伯基特淋巴瘤细胞系中的肿瘤B细胞以及血液样本中的健康T和B淋巴细胞组成的异质人活淋巴细胞混合物。GrFFF不需要使用荧光免疫标签来分选细胞，并且可以收集分选后的细胞进行进一步表征。使用流式细胞术评估收集到的细胞的活力，并评估实现的细胞分离效果。在通过GrFFF获得的特定组分中发现了少量肿瘤B淋巴细胞（少于2%）。克隆形成试验证实了肿瘤细胞的高去除率（超过98%）。
