Dumont Courtney M, Karande Pankaj, Thompson Deanna M
1 Department of Biomedical Engineering, Rensselaer Polytechnic Institute , Troy, New York.
Tissue Eng Part C Methods. 2014 Aug;20(8):620-9. doi: 10.1089/ten.TEC.2013.0362. Epub 2014 Mar 31.
Designing an ideal biomaterial supportive of multicellular tissue repair is challenging, especially with a poor understanding of the synergy between constituent proteins and growth factors. A brute-force approach, based on screening all possible combinations of proteins and growth factors, is inadequate due to the prohibitively large experimental space coupled with current low-throughput screening techniques. A high-throughput screening platform based on rational and combinatorial strategies for design and testing of proteins and growth factors can significantly impact the discovery of novel tissue-specific biomaterials. Here, we report the development of a flexible high-throughput screening platform, Rapid Assessment of Migration and Proliferation (RAMP), to rapidly investigate cell viability, proliferation, and migration in response to highly miniaturized three-dimensional biomaterial cultures (4-20 μL) with sparingly low cell densities (63-1000 cells per μL for cell arrays; 1 μL of 1000-10,000 cells per μL for migration arrays). The predictions made by RAMP on the efficacy and potency of the biomaterials are in agreement with the predictions made by conventional assays but at a throughput that is at least 100-1000-fold higher. The RAMP assay is therefore a novel approach for the rapid discovery of tissue-specific biomaterials for tissue engineering and regenerative medicine.
设计一种理想的支持多细胞组织修复的生物材料具有挑战性,尤其是在对组成蛋白质和生长因子之间的协同作用了解不足的情况下。基于筛选蛋白质和生长因子的所有可能组合的蛮力方法是不够的,因为实验空间过大,加上当前的低通量筛选技术。基于合理和组合策略的用于蛋白质和生长因子设计与测试的高通量筛选平台,可能会对新型组织特异性生物材料的发现产生重大影响。在此,我们报告了一种灵活的高通量筛选平台——迁移和增殖快速评估(RAMP)的开发,以快速研究细胞在响应高度微型化的三维生物材料培养物(4 - 20 μL)时的活力、增殖和迁移情况,这些培养物具有极低的细胞密度(细胞阵列每微升63 - 1000个细胞;迁移阵列每微升1 μL含1000 - 10,000个细胞)。RAMP对生物材料功效和效力的预测与传统检测方法的预测一致,但通量至少高100 - 1000倍。因此,RAMP检测是一种用于快速发现用于组织工程和再生医学的组织特异性生物材料的新方法。