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International Journal of Bioprinting Kidney hydrogel print for renal cancer model

Figure 4. Biocompatibility tests of GelMA-Kidney hydrogel. (A) ACHN cellular morphology; (B) ACHN cellular proliferation; (C–L) Expression levels of
an array of EMT-related markers in ACHN cells in various concentrations of GelMA-Kidney hydrogels.

Next, Kyoto Encyclopedia of Genes and Genomes compared to the 10% GelMA control, while CDH1 was
(KEGG) enrichment analysis was also conducted to downregulated. This observation was further verified
analyze changes in signaling pathways of ACHN cells by the quantification of immunofluorescence intensity
grown in 10% GelMA-2% Kidney cultures (Figure 5C). using NIH ImageJ (Figure 6B–F). Here, the expression
We identified several signaling pathways related to cancer of vimentin, TWIST, CD44, CDH1, and CDH2 was
development, including the VEGF (hsa04370), PPAR up- or downregulated by 2.53-, 1.52-, 2.00-, 0.71-, and
(hsa03320), PI3K-Akt (hsa04151), HIF-1 (hsa04066), and 2.11-folds, respectively.
TGF-beta (hsa04350).
3.6. ACHN drug resistance
3.5. Expression of EMT-related protein Finally, the effect of dECM powders on ACHN drug
To examine the effect of kidney dECM powder on resistance was evaluated by assessing the resistance of the
ACHN behavior, we evaluated the effect of the 10%
GelMA-2% Kidney on EMT-related protein expression. cells toward sunitinib on day 5 (Figure S2 in Supplementary
Specifically, we tested for vimentin, TWIST, CD44, File). The cellular viabilities of the 2D, 10% GelMA, and
CDH1, and CDH2, by immunofluorescence staining on 10% GelMA-2% Kidney cultures were 36.40% ± 1.08%,
day 5 of the 10% GelMA-2% Kidney treatment (Figure 40.19% ± 7.02%, and 46.41% ± 4.93%, respectively. ACHN
6A). The result for secondary antibody assay is shown cells cultured in GelMA-based hydrogels all showed
in Figure S2K (Supplementary File), where no positive significantly higher chemoresistance compared with 2D
staining was observed, indicating the genuine positivity cell cultures. Moreover, the inclusion of 2% kidney dECM
in the fluorescence images. We found that the protein powder significantly promoted cellular viability compared
expression of vimentin, TWIST, CD44, and CDH2 was to 10% GelMA, suggesting that kidney dECM powder can
upregulated in the 10% GelMA-2% Kidney culture enhance the drug resistance of ACHN cells.

Volume 10 Issue 2 (2024) 288 doi: 10.36922/ijb.1413