Patterned PVA Hydrogels with 3D Petri Dish<sup>®</sup> Micro-Molds of Varying Topography for Spheroid Formation of HeLa Cancer Cells: In Vitro Assessment
Maira Moreno Valtierra,
Adriana Urue Corral,
Jorge Armando Jiménez-Avalos,
Erika Barbosa Avalos,
Judith Dávila-Rodríguez,
Norma Morales Hernández,
Mauricio Comas-García,
Guillermo Toriz González,
Antonio Oceguera-Villanueva,
José Alfonso Cruz-Ramos,
Rodolfo Hernández Gutiérrez,
Moisés Martínez Velázquez,
Zaira Yunuen García Carvajal
Affiliations
Maira Moreno Valtierra
Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco (CIATEJ), Av. Normalistas # 800, Col. Colinas de la Normal, Guadalajara 44270, Mexico
Adriana Urue Corral
Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco (CIATEJ), Av. Normalistas # 800, Col. Colinas de la Normal, Guadalajara 44270, Mexico
Jorge Armando Jiménez-Avalos
Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco (CIATEJ), Av. Normalistas # 800, Col. Colinas de la Normal, Guadalajara 44270, Mexico
Erika Barbosa Avalos
Laboratorio de Anatomía Patológica, Hospital Civil Viejo Fray Antonio Alcalde, Coronel Calderón #777, El Retiro, Guadalajara 44280, Mexico
Judith Dávila-Rodríguez
Laboratorio de Anatomía Patológica, Hospital Civil Viejo Fray Antonio Alcalde, Coronel Calderón #777, El Retiro, Guadalajara 44280, Mexico
Norma Morales Hernández
Tecnología Alimentaria, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco (CIATEJ), Camino Arenero # 1227, Col. El Bajío del Arenal, Zapopan 45019, Mexico
Mauricio Comas-García
Facultad de Ciencias, Universidad Autónoma de San Luis Potosí, Av. Parque Chapultepec # 1570, San Luis Potosí 78210, Mexico
Guillermo Toriz González
Departamento de Madera, Celulosa y Papel, Centro Universitario de Ciencias Exactas e Ingenierías, Universidad de Guadalajara, Carretera Guadalajara-Nogales km 15.5, Zapopan 45220, Mexico
Antonio Oceguera-Villanueva
Instituto Jalisciense de Cancerología, Secretaría de Salud Jalisco, 715 Coronel Calderón St., El Retiro, Guadalajara 44280, Mexico
José Alfonso Cruz-Ramos
Instituto Jalisciense de Cancerología, Secretaría de Salud Jalisco, 715 Coronel Calderón St., El Retiro, Guadalajara 44280, Mexico
Rodolfo Hernández Gutiérrez
Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco (CIATEJ), Av. Normalistas # 800, Col. Colinas de la Normal, Guadalajara 44270, Mexico
Moisés Martínez Velázquez
Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco (CIATEJ), Av. Normalistas # 800, Col. Colinas de la Normal, Guadalajara 44270, Mexico
Zaira Yunuen García Carvajal
Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco (CIATEJ), Av. Normalistas # 800, Col. Colinas de la Normal, Guadalajara 44270, Mexico
Cell spheroids are an important three-dimensional (3D) model for in vitro testing and are gaining interest for their use in clinical applications. More natural 3D cell culture environments that support cell–cell interactions have been created for cancer drug discovery and therapy applications, such as the scaffold-free 3D Petri Dish® technology. This technology uses reusable and autoclavable silicone micro-molds with different topographies, and it conventionally uses gelled agarose for hydrogel formation to preserve the topography of the selected micro-mold. The present study investigated the feasibility of using a patterned Poly(vinyl alcohol) hydrogel using the circular topography 12–81 (9 × 9 wells) micro-mold to form HeLa cancer cell spheroids and compare them with the formed spheroids using agarose hydrogels. PVA hydrogels showed a slightly softer, springier, and stickier texture than agarose hydrogels. After preparation, Fourier transform infrared (FTIR) spectra showed chemical interactions through hydrogen bonding in the PVA and agarose hydrogels. Both types of hydrogels favor the formation of large HeLa spheroids with an average diameter of around 700–800 µm after 72 h. However, the PVA spheroids are more compact than those from agarose, suggesting a potential influence of micro-mold surface chemistry on cell behavior and spheroid formation. This was additionally confirmed by evaluating the spheroid size, morphology, integrity, as well as E-cadherin and Ki67 expression. The results suggest that PVA promotes stronger cell-to-cell interactions in the spheroids. Even the integrity of PVA spheroids was maintained after exposure to the drug cisplatin. In conclusion, the patterned PVA hydrogels were successfully prepared using the 3D Petri Dish® micro-molds, and they could be used as suitable platforms for studying cell–cell interactions in cancer drug therapy.
