Ecole Normale Supérieure de Cachan, CNRS, SATIE, UMR 8029, Cachan, France ; Univ Paris-Sud, CNRS, Institut d'Electronique Fondamentale, UMR 8622, Orsay, France.
Ecole Normale Supérieure de Cachan, CNRS, SATIE, UMR 8029, Cachan, France.
Biomicrofluidics. 2013 Jul 3;7(4):44101. doi: 10.1063/1.4813062. eCollection 2013.
Cell fusion consists of inducing the formation of a hybridoma cell containing the genetic properties of the progenitor cells. Such an operation is usually performed chemically or electrically. The latter method, named electrofusion, is considered as having a strong potential, due to its efficiency and non-toxicity, but deserves further investigations prior to being applicable for key applications like antibody production and cancer immunotherapy. Indeed, to envision such applications, a high amount of hybrid cells is needed. In this context, we present in this paper a device for massive cell pairing and electrofusion, using a microarray of non-connected conductive pads. The electrofusion chamber--or channel--exposes cells to an inhomogeneous electric field, caused by the pads array, enabling the trapping and pairing of cells with dielectrophoresis (DEP) forces prior to electrofusion. Compared to a mechanical trapping, such electric trapping is fully reversible (on/off handling). The DEP force is contactless and thus eases the release of the produced hybridoma. Moreover, the absence of wire connections on the pads permits the high density trapping and electrofusion of cells. In this paper, the electric field mapping, the effect of metallic pads thickness, and the transmembrane potential of cells are studied based on a numerical model to optimize the device. Electric calculations and experiments were conducted to evaluate the trapping force. The structure was finally validated for cell pairing and electrofusion of arrays of cells. We believe that our approach of fully electric trapping with a simple structure is a promising method for massive production of electrofused hybridoma.
细胞融合包括诱导形成含有亲本细胞遗传特性的杂交瘤细胞。这种操作通常通过化学或电方法进行。后一种方法称为电融合，由于其效率高、毒性低，被认为具有很大的潜力，但在适用于抗体生产和癌症免疫治疗等关键应用之前，还需要进一步研究。事实上，为了实现这些应用，需要大量的杂交细胞。在这方面，我们在本文中提出了一种使用非连接导电垫微阵列进行大规模细胞配对和电融合的装置。电融合室——或通道——使细胞暴露于由垫阵列引起的非均匀电场中，从而能够在电融合之前利用介电泳（DEP）力捕获和配对细胞。与机械捕获相比，这种电捕获是完全可逆的（开/关处理）。DEP 力是无接触的，因此便于释放产生的杂交瘤。此外，垫上没有金属线连接，允许高密度捕获和电融合细胞。本文基于数值模型研究了电场映射、金属垫厚度的影响以及细胞的跨膜电位，以优化器件。进行了电计算和实验以评估捕获力。最后，该结构经过验证可用于细胞对和细胞阵列的电融合。我们相信，我们的这种具有简单结构的全电捕获方法是大规模生产电融合杂交瘤的有前途的方法。
