Wang Yuli, Phillips Colleen N, Herrera Gabriela S, Sims Christopher E, Yeh Jen Jen, Allbritton Nancy L
Department of Chemistry, University of North Carolina, Chapel Hill, NC 27599.
RSC Adv. 2013 Jun 28;3(24):9264-9272. doi: 10.1039/C3RA41764F.
A new strategy for efficient sorting and implantation of viable adherent cells into animals is described. An array of biodegradable micro-structures (microrafts) was fabricated using a polydimethylsiloxane substrate for micromolding poly(lactic-co-glycolic acid) (PLGA). Screening various forms of PLGA determined that the suitability of PLGA for microraft manufacture, biocompatibility and degradation was dependent on molecular weight and lactic/glycolic ratio. Cells plated on the array selectively attached to the microrafts and could be identified by their fluorescence, morphology or other criteria. The cells were efficiently dislodged and collected from the array using a microneedle device. The platform was used to isolate specific cells from a mixed population establishing the ability to sort target cells for direct implantation. As a proof of concept, fluorescently conjugated microrafts carrying tumor cells stably expressing luciferase were isolated from an array and implanted subcutaneously into mice. bio-luminescence imaging confirmed the growth of a tumor in the recipient animals. Imaging of tissue sections from the tumors demonstrated degradation of the implanted microrafts. The process is a new strategy for isolating and delivering a small number of adherent cells for animal implantation with potential applications in tissue repair, tumor induction, differentiation of stem cells and other biomedical research.
本文描述了一种将活的贴壁细胞高效分选并植入动物体内的新策略。使用聚二甲基硅氧烷底物进行聚乳酸-乙醇酸共聚物(PLGA)微成型,制造了一系列可生物降解的微结构(微筏)。对各种形式的PLGA进行筛选后确定,PLGA用于微筏制造、生物相容性和降解的适用性取决于分子量和乳酸/乙醇酸比例。接种在阵列上的细胞选择性地附着在微筏上,并可通过其荧光、形态或其他标准进行识别。使用微针装置可有效地从阵列上分离并收集细胞。该平台用于从混合群体中分离特定细胞,确立了分选靶细胞以进行直接植入的能力。作为概念验证,从阵列中分离出携带稳定表达荧光素酶的肿瘤细胞的荧光共轭微筏,并将其皮下植入小鼠体内。生物发光成像证实了受体动物体内肿瘤的生长。对肿瘤组织切片的成像显示植入的微筏发生了降解。该过程是一种分离和递送少量贴壁细胞用于动物植入的新策略,在组织修复、肿瘤诱导、干细胞分化及其他生物医学研究中具有潜在应用价值。