A novel 3D biofabrication strategy to improve cell proliferation and differentiation of human Wharton’s jelly mesenchymal stromal cells for cell therapy and tissue engineering
MetadataShow full item record
AuthorBlanco Elices, Cristina; Sánchez Porras, David; Chato Astrain, Jesús; Campos Sánchez, Fernando; Alaminos Mingorance, Miguel; Garzón Bello, Ingrid Johanna; Campos Muñoz, Antonio Jesús
Cell cultureThree-dimensionalBiofabricationHuman Wharton’s jelly mesenchymal stromal cellsCell therapyTissue engineering
Blanco-Elices C, Oruezabal RI, Sánchez-Porras D, Chato-Astrain J, Campos F, Alaminos M, Garzón I and Campos A (2023). A novel 3D biofabrication strategy to improve cell proliferation and differentiation of human Wharton’s jelly mesenchymal stromal cells for cell therapy and tissue engineering. Front. Bioeng. Biotechnol. 11:1235161. [doi: 10.3389/fbioe.2023.1235161]
SponsorshipUniversidad de Granada, Spain; Consejería de Transformación Económica, Industria, Conocimiento y Universidades; FEDER PE-0395-2019; Secretaría de Estado de Investigación, Desarrollo e Innovación FIS PI18/0331, FIS PI20/0317, FIS PI21/0980, FIS PI22/0059 I+D+i; Instituto de Salud Carlos III ISCIII; Ministerio de Ciencia e Innovación MICINN; European Regional Development Fund ERDF; Consejería de Salud y Familias, Junta de Andalucía B-CTS-450-UGR20; Spanish National Plan for Scientific and Technical Research and Innovation
Purpose: Obtaining sufficient numbers of cells in a short time is a major goal of cell culturing in cell therapy and tissue engineering. However, current bidimensional (2D) culture methods are associated to several limitations, including low efficiency and the loss of key cell differentiation markers on cultured cells. Methods: In the present work, we have designed a novel biofabrication method based on a three-dimensional (3D) culture system (FIBRIAGAR-3D). Human Wharton’s jelly mesenchymal stromal cells (HWJSC) were cultured in 3D using 100%, 75%, 50%, and 25% concentrations of fibrin-agarose biomaterials (FA100, FA75, FA50 and FA25 group) and compared with control cells cultured using classical 2D systems (CTR-2D). Results: Our results showed a significant increase in the number of cells generated after 7 days of culture, with cells displaying numerous expansions towards the biomaterial, and a significant overexpression of the cell proliferation marker KI67 was found for the FA75 and FA100 groups. TUNEL and qRT-PCR analyses demonstrated that the use of FIBRIAGAR-3D was not associated with an induction of apoptosis by cultured cells. Instead, the 3D system retained the expression of typical phenotypic markers of HWJSC, including CD73, CD90, CD105, NANOG and OCT4, and biosynthesis markers such as types-I and IV collagens, with significant increase of some of these markers, especially in the FA100 group. Finally, our analysis of 8 cell signaling molecules revealed a significant decrease of GM-CSF, IFN-g, IL2, IL4, IL6, IL8, and TNFα, suggesting that the 3D culture system did not induce the expression of pro-inflammatory molecules. Conclusion: These results confirm the usefulness of FIBRIAGAR-3D culture systems to increase cell proliferation without altering cell phenotype of immunogenicity and opens the door to the possibility of using this novel biofabrication method in cell therapy and tissue engineering of the human cornea, oral mucosa, skin, urethra, among other structures.