Seymour Samantha, Cadena Ines, Johnson Mackenzie, Thakkar Riya, Jenne Molly, Adem Iman, Almer Alyssa, Frankovic Rachael, Spence Danielle, Haddadin Andrea, Fogg Kaitlin C
School of Chemical, Biological, and Environmental Engineering, Oregon State University, Corvallis, OR 97330 USA.
HP Inc., Corvallis, OR USA.
Cell Mol Bioeng. 2025 Jan 23;18(1):71-82. doi: 10.1007/s12195-024-00841-y. eCollection 2025 Feb.
Cervical and endometrial cancers pose significant challenges in women's healthcare due to their high mortality rates and limited treatment options. High throughput screening (HTS) of cervical and endometrial cancer in vitro models offers a promising avenue for drug repurposing and broadening patient treatment options. Traditional two-dimensional (2D) cell-based screenings have limited capabilities to capture crucial multicellular interactions, that are improved upon in three dimensional (3D) multicellular tissue engineered models. However, manual fabrication of the 3D platforms is both time consuming and subject to variability. Thus, the goal of this study was to utilize automated cell dispensing to fabricate 3D cell-based HTS platforms using the HP D100 Single Cell Dispenser to dispense cervical and endometrial cancer cells.
We evaluated the effects of automated dispensing of the cancer cell lines by tuning the dispensing protocol to align with cell size measured in solution and the minimum cell number for acceptable cell viability and proliferation. We modified our previously reported coculture models of cervical and endometrial cancer to be in a 384 well plate format and measured microvessel length and cancer cell invasion.
Automatically and manually dispensed cells were directly compared revealing minimal differences between the dispensing methods. These findings suggest that automated dispensing of cancer cells minimally affects cell behavior and can be deployed to decrease in vitro model fabrication time.
By streamlining the manufacturing process, automated dispensing holds promise for enhancing efficiency and scalability of 3D in vitro HTS platforms, ultimately contributing to advancement in cancer research and treatment.
The online version contains supplementary material available at 10.1007/s12195-024-00841-y.
宫颈癌和子宫内膜癌因其高死亡率和有限的治疗选择，在女性医疗保健中构成重大挑战。对宫颈癌和子宫内膜癌体外模型进行高通量筛选（HTS）为药物再利用和拓宽患者治疗选择提供了一条有前景的途径。传统的基于二维（2D）细胞的筛选在捕捉关键的多细胞相互作用方面能力有限，而三维（3D）多细胞组织工程模型则有所改进。然而，3D平台的手工制作既耗时又存在变异性。因此，本研究的目标是利用自动细胞分配技术，使用惠普D100单细胞分配器来分配宫颈癌细胞和子宫内膜癌细胞，从而构建基于3D细胞的HTS平台。
我们通过调整分配方案，使其与溶液中测量的细胞大小以及可接受的细胞活力和增殖所需的最小细胞数量相匹配，来评估癌细胞系自动分配的效果。我们将之前报道的宫颈癌和子宫内膜癌共培养模型修改为384孔板形式，并测量微血管长度和癌细胞侵袭情况。
对自动分配和手动分配的细胞进行了直接比较，发现分配方法之间的差异极小。这些发现表明，癌细胞的自动分配对细胞行为的影响最小，并且可以用于减少体外模型的制作时间。
通过简化制造过程，自动分配有望提高基于3D的体外HTS平台的效率和可扩展性，最终推动癌症研究和治疗的进展。
在线版本包含可在10.1007/s12195-024-00841-y获取的补充材料。
