3D Bioprinted Senescent Caco2 Modeling Using a Low-Cost DIY 3D Bioprinter: Challenges and Limitations
Sofian Al Shboul, Yazan Al Dweiri, Mai F. AlSakhen, Nisreen Himsawi, Helen Al Zain, Sabal Al Hadidi, Raghda Barham, Nidaa A. Ababneh, Ashraf I. Khasawneh, Moureq R. Alotaibi, Lubna H. Tahtamouni, Tareq SalehBackground:
Therapy-induced senescence (TIS) is a major component of tumor cell response to a variety of anticancer treatments in preclinical models. Most TIS-related literature is derived from 2D-based studies with limited evidence in 3D models. This study aimed to investigate the challenges of developing an in-house (DIY) 3D bioprinted TIS cell model.
Materials and Methods:
A conventional thermoplastic extrusion-based 3D printer (Anet A8 kit) was modified to function as a bioprinter, enabling the fabrication of a Caco2 cell model via extrusion-based bioprinting. The printed 3D models were exposed to doxorubicin (DOX, at 1 and 10 μM), a well-established TIS trigger, and then assessed for senescence-associated β-galactosidase (SA-β-gal) activity and gene expression of several senescence regulators and components of the senescence-associated secretory phenotype (SASP) via real-time PCR.
Results:
Treated cells exhibited increased SA-β-gal staining and increased expression of CDKN2A and CDKN2B (and modestly TP53). Furthermore, the expression of SASP-related genes, including MMP3, MMP9, CXCL8, and TGF-β1 (but not IL-1A), was increased, al-beit non-significantly, suggesting a potential opportunity to detect transcriptional SASP changes in 3D bioprinted tumor cell models.
Discussion:
This study provides an early attempt to develop a TIS 3D bioprinted tumor cell model, highlighting several challenges, including infrequent nozzle clogging, difficulty of image-based quantification of SA-β-gal staining, and inter-print variation in gene expression profiles.
Conclusion:
In-house DIY 3D bioprinting for investigating tumor cell senescence is feasible but requires further improvement prior to use in senolytic screening.