Cell seeding simulation on micropatterned islands shows cell density depends on area to perimeter ratio, not on island size or shape.,Acta Biomaterialia

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Cell seeding simulation on micropatterned islands shows cell density depends on area to perimeter ratio, not on island size or shape.
Acta Biomaterialia ( IF 9.4 ) Pub Date : 2020-02-27 , DOI: 10.1016/j.actbio.2020.02.035
Zachary T Berent ₁ , Amy J Wagoner Johnson ₂

Affiliation

Protein micropatterned substrates have been used to control cell size, shape, and cell-cell contacts, characteristics that influence a range of cell behaviors from early cell differentiation to late stages of maturation. Knowing the initial island cell seeding density is important to interpreting results and understanding downstream cell behavior. While studies routinely report the intended or target cell seeding density, they do not report the actual cell seeding density on the islands. As cells proliferate, differences in initial cell seeding density could compound and may lead to misinterpretation of results. In this work, we present a cell seeding simulation and apply it to 100s of islands with a range of geometries (sizes and shapes) to explore how island cell seeding density relates to the target or unpatterned cell seeding density. We first experimentally validate the simulation and then show that normalized island cell seeding density depends on island size, shape, and spacing, but can be predicted solely from island area to perimeter ratio, A2P, via a power law relationship for a wide range of island geometries. Interestingly, normalized island cell seeding density is the same as the normalized unpatterned cell seeding density for A2P ≥ 17 µm. This simulation will help to design micropatterned substrates and to have more accurate representation of the island cell seeding density at the start of experiments. By knowing the island cell seeding density, we can more easily reproduce results across research groups to understand the roles of cell-cell contact and cell size and shape on cell behavior. STATEMENT OF SIGNIFICANCE: We present a cell seeding simulation on protein-micropatterned substrates and use it to simulate seeding across 100s of island geometries (size, shape, and spacing) covering two orders of magnitude in size. The simulation shows that island cell density varies significantly with island geometry compared to the target seeding density. However, island cell density can be predicted from one geometric parameter - the island's area to perimeter ratio. Results will help direct researchers on how to achieve uniform cell density across all island geometries. Since cell density and island shape both influence cell behaviors, such as differentiation, this simulation may help to isolate these factors, facilitate micropatterned substrate design, and provide a mechanism for more reproduceable results across studies.

中文翻译：

在微图案岛上进行的细胞播种模拟显示，细胞密度取决于面积与周长的比率，而不取决于岛的大小或形状。

蛋白质微图案化的底物已用于控制细胞大小，形状和细胞间接触，这些特征会影响从早期细胞分化到成熟后期的一系列细胞行为。了解初始岛细胞接种密度对于解释结果和理解下游细胞行为很重要。尽管研究常规报告了预期或目标的细胞接种密度，但并未报告岛上的实际细胞接种密度。随着细胞的增殖，初始细胞接种密度的差异可能会加剧，并可能导致对结果的错误理解。在这项工作中，我们提出了一个细胞播种模拟，并将其应用于具有各种几何形状（大小和形状）的100个岛上，以探索岛细胞播种密度与目标或无图案细胞播种密度之间的关系。我们首先通过实验验证仿真结果，然后表明归一化的岛细胞接种密度取决于岛的大小，形状和间距，但可以通过幂律关系从大范围的岛上单独从岛面积与周长比A2P进行预测几何形状。有趣的是，归一化的岛细胞接种密度与A2P≥17 µm的归一化无图案细胞接种密度相同。此模拟将有助于设计微图案化的底物，并在实验开始时更准确地表示岛细胞的接种密度。通过了解岛细胞的接种密度，我们可以更轻松地在各个研究小组之间复制结果，以了解细胞与细胞的接触以及细胞大小和形状对细胞行为的作用。重要性声明：我们介绍了一种在蛋白质微图案化基质上进行的细胞播种模拟，并使用它来模拟跨越100个岛的几何形状（大小，形状和间距）的播种，覆盖了两个数量级的大小。仿真显示，与目标播种密度相比，岛细胞密度随岛的几何形状显着变化。但是，可以从一个几何参数-岛屿的面积与周长之比来预测岛屿的细胞密度。研究结果将指导研究人员如何在所有岛上实现均匀的细胞密度。由于细胞密度和岛状形状均会影响细胞行为，例如分化，因此该模拟可能有助于分离这些因素，促进微图案化底物设计，并为整个研究提供更可再现的结果的机制。

更新日期：2020-02-27

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