(B-61) Dispensing Gynecological Cancer Cells to Fabricate 3D Cervical and Endometrial Cancer Platforms

Introduction:: There is a critical need for cancer models that capture the tumor microenvironment and can interface with high throughput drug screening methods to assess anti-metastatic and anti-angiogenic drug efficacy in a rapid yet high content manner. Our lab has developed a novel multilayer multicellular platform of cancer cells and endothelial cells that is compatible with standard high throughput screening methods. Specifically, we developed novel three-dimensional models of cervical and endometrial cancer in 384-well plate formats that maximize phenotypic response: cancer growth, cancer invasion, and endothelial microvessel formation simultaneously as these are all potential targets for anti-cancer therapies (Figure 1A). Our multilayer multicellular models must be automatically reproduced to perform large-scale compound screening to identify new treatment options for cervical and endometrial cancers from the FDA approved drug library.

Materials and Methods:: Human cervical cancer cells (SiHa, Ca Ski) and human endometrial cancer cells (HEC-1A) were dispensed using the HP D100 Single Cell Dispenser on tissue culture plastic, gelatin methacryloyl (GelMA) based hydrogels, and Matrigel. Human microvascular endothelial cells (hMVECs) and human umbilical vein endothelial cells (hUVECs) were manually dispensed on GelMA and polyethylene glycol diacrylate (PEGDA) based hydrogels. The hydrogel layers fabricated for the multilayer multicellular models were composed of optimized concentrations of collagen type I and IV, fibrinogen, thrombin, fibronectin, laminin, GelMA, and PEGDA. These gel compositions have been optimized for each layer of the cervical cancer and endometrial cancer model to maximize phenotypic response. The automatically dispensed (AD) cancer cells of the multilayer multicellular models were compared to manually dispensed cancer cells (MD) under the same conditions.

Results, Conclusions, and Discussions:: Results: Dispensing method precision for automatically dispensed (AD) and manually dispensed cells (MD) was evaluated by comparing the target number of cells dispensed per well to the observed number of cells with three biological replicates of each cell type: SiHa, Ca Ski, HEC-1A (Figure 1B). There was no significant difference between the AD replicates, with p < 0.12. In contrast, the MD replicates had significant differences between replicates, with p < 0.0001. There were no significant differences in viability across all cell lines examined (Figure 1C). Additionally, there were no significant differences observed in cell proliferation, viability, micro-vessel length, or cancer invasion depth between the MD and AD cancer cells (Figure 1D). This was true for both the cervical cancer cell lines on the cervical cancer model and the endometrial cancer cell line on the endometrial cancer model.

Conclusion: There were minimal effects on cell proliferation, viability, and phenotypic response when automatically dispensing cervical and endometrial cancer cells using the HP D100 Single Cell Dispenser. Additionally, increased reproducibility is observed with automatic dispensing compared to manual dispensing. Automizing the fabrication of these multilayer multicellular models enables our team to perform large-scale hit identification studies to increase treatment options and improve personalized medicine for cervical and endometrial cancer patients.


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