A decellularized extracellular matrix hydrogel to promote the proliferation of human salivary gland cells
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Abstract
Objectives: Matrigel, a raw matrix from the Engelbreth-Holm-Swarm mouse sarcoma, has been commonly used to promote salivary gland (SG) cell assembly and proliferation in vitro, however, it possesses limitations such as batch-to-batch variations and undefined tumor-derived components, hence, alternative matrices are lacking. This study aimed to develop porcine submandibular gland decellularized extracellular matrix (SG-dECM) hydrogels to support SG cell viability and proliferation.
Materials and methods: SG-dECM was produced using non-ionic and ionic detergent perfusions, then digested with a pepsin-based HCl buffer to generate SG-dECM hydrogels at concentrations of 1, 5, and 10 mg/mL. SG-dECM sections were stained with hematoxylin and eosin (H&E), and rhodamine-labeled peanut agglutinin staining of glycoproteins/mucins. A human submandibular gland cell line, A253 (HTB-41TM, ATCC), was cultured as a monolayer culture with SG-dECM hydrogel- or Matrigel-coated on 96-well plates and assessed for proliferation over 4 days using an ATP-dependent assay. Uncoated wells were used as negative controls. Viable and late apoptotic cells were quantified with calcein-AM and propidium iodide staining, respectively. One- and two-way ANOVA with Tukey’s post-hoc tests were performed with 5 biological replicates.
Results: The developed SG-dECM retained moderately glycoproteins and mucins while cellular components were effectively removed. SG-dECM hydrogels at 5 mg/mL significantly enhanced A253 cell proliferation and viability compared to Matrigel (p<0.05) and uncoated wells (p<0.001) after 4 culture days.
Conclusion: SG-dECM hydrogels at 5 mg/mL promoted the proliferation and viability of A253 cells over 4 culture days. Thus, SG-dECM hydrogel could be a viable alternative to Matrigel for future drug screening applications.
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