Cell Seeding Technology for Microarray-Based Quantitative Human Primary Skeletal Muscle Cell Analysis,Analytical Chemistry

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Cell Seeding Technology for Microarray-Based Quantitative Human Primary Skeletal Muscle Cell Analysis
Analytical Chemistry ( IF 6.7 ) Pub Date : 2019-11-01 , DOI: 10.1021/acs.analchem.9b03722
Hyun Young Shin , Seung Joon Lee , Hyung Woo Seo , Min Young Kim , Aseer Intisar , Kyungmoo Yea , Sung Chun Cho , Yun-Il Lee , Young Zoon Kim ₁ , Ogan Gurel , Danny van Noort ₂ , Sang Chul Park , Minseok S. Kim

Affiliation

Pipetting techniques play a crucial role in obtaining reproducible and reliable results, especially when seeding cells on small target areas, such as on microarrays, biochips or microfabricated cell culture systems. For very rare cells, such as human primary skeletal muscle cells (skMCs), manual (freehand) cell seeding techniques invariably result in nonuniform cell spreading and heterogeneous cell densities, giving rise to undesirable variations in myogenesis and differentiation. To prevent such technique-dependent variation, we have designed and fabricated a simple, low-cost pipet guidance device (PGD), and holder that works with hand-held pipettes. This work validates the accuracy and reproducibility of the PGD platform and compares its effectiveness with manual and robotic seeding techniques. The PGD system ensures reproducibility of cell seeding, comparable to that of more expensive robotic dispensing systems, resulting in a high degree of cell uniformity and homogeneous cell densities, while also enabling cell community studies. As compared to freehand pipetting, PGD-assisted seeding of C2C12 mouse myoblasts showed 5.3 times more myotube formation and likewise myotubes derived from PGD-seeded human primary skMCs were 3.6 times thicker and 2.2 times longer. These results show that this novel, yet simple PGD-assisted pipetting technique provides precise cell seeding on small targets, ensuring reproducible and reliable high-throughput cell assays.

中文翻译：

基于微阵列的定量人类原代骨骼肌细胞分析的细胞接种技术

移液技术在获得可再现和可靠的结果中起着至关重要的作用，尤其是在将细胞播种在较小目标区域（例如微阵列，生物芯片或微细化细胞培养系统）上时。对于非常稀有的细胞，例如人原代骨骼肌细胞（skMCs），手工（徒手）细胞接种技术总是会导致细胞扩散不均匀和细胞密度异质，从而在成肌和分化过程中引起不良变化。为了防止这种依赖技术的变化，我们设计并制造了一种简单，低成本的移液器导引装置（PGD）和可与手持式移液器配合使用的支架。这项工作验证了PGD平台的准确性和可重复性，并将其有效性与手动和自动播种技术进行了比较。与更昂贵的自动分配系统相比，PGD系统可确保细胞接种的可重复性，从而实现高度的细胞均匀性和均一的细胞密度，同时还能够进行细胞群落研究。与徒手移液相比，PGD辅助播种的C2C12小鼠成肌细胞显示出5.3倍以上的肌管形成，同样，源自PGD播种的人原发性skMC的肌管厚3.6倍，长2.2倍。这些结果表明，这种新颖而又简单的PGD辅助移液技术可在小靶标上提供精确的细胞接种，从而确保可重现和可靠的高通量细胞测定。与徒手移液相比，PGD辅助播种的C2C12小鼠成肌细胞显示出5.3倍以上的肌管形成，同样，源自PGD播种的人原发性skMC的肌管厚3.6倍，长2.2倍。这些结果表明，这种新颖而又简单的PGD辅助移液技术可在小靶标上提供精确的细胞接种，从而确保可重现和可靠的高通量细胞测定。与徒手移液相比，PGD辅助播种的C2C12小鼠成肌细胞显示出5.3倍以上的肌管形成，同样，源自PGD播种的人原发性skMC的肌管厚3.6倍，长2.2倍。这些结果表明，这种新颖而又简单的PGD辅助移液技术可在小靶标上提供精确的细胞接种，从而确保可重现和可靠的高通量细胞测定。

更新日期：2019-11-01

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