The Use of Nanofibrillar Cellulose Hydrogel As a Flexible Three-Dimensional Model to Culture Human Pluripotent Stem Cells

Abstract

Human embryonic stem cells and induced pluripotent stem cells have great potential in research and therapies. The current in vitro culture systems for human pluripotent stem cells (hPSCs) do not mimic the threedimensional (3D) in vivo stem cell niche that transiently supports stem cell proliferation and is subject to changes which facilitate subsequent differentiation during development. Here, we demonstrate, for the first time, that a novel plant-derived nanofibrillar cellulose (NFC) hydrogel creates a flexible 3D environment for hPSC culture. The pluripotency of hPSCs cultured in the NFC hydrogel was maintained for 26 days as evidenced by the expression of OCT4, NANOG, and SSEA-4, in vitro embryoid body formation and in vivo teratoma formation. The use of a cellulose enzyme, cellulase, enables easy cell propagation in 3D culture as well as a shift between 3D and two-dimensional cultures. More importantly, the removal of the NFC hydrogel facilitates differentiation while retaining 3D cell organization. Thus, the NFC hydrogel represents a flexible, xeno-free 3D culture system that supports pluripotency and will be useful in hPSC-based drug research and regenerative medicine.

Producing ultrapure wood cellulose nanofibrils and evaluating the cytotoxicity using human skin cells

“…While CNF materials have been compatible with different cell lines (no cytotoxicity) (Alexandrescu, Syverud, Gatti & Chinga-Carrasco, 2013;Bhattacharya et al, 2012;Hua et al, 2014;Lou et al, 2014;Malinen et al, 2014;Tehrani, Nordli, Pukstad, Gethin & Chinga-Carrasco;Vartiainen et al, 2011), a concentration dependent reduction in metabolic activity and/or cell proliferation has also been seen (Čolić, Mihajlović, Mathew, Naseri & Kokol, 2015). TEMPO-mediated oxidation and carboxymethylation are relatively common pre-treatments applied to facilitate the deconstruction of the fibre cell wall and thus the production of CNF (Saito, Nishiyama, Putaux, Vignon & Isogai, 2006;Wågberg, Decher, Norgren, Lindström, Ankerfors & Axnäs, 2008).…”

Internalization of (bis)phosphonate-modified cellulose nanocrystals by human osteoblast cells

“…Nanocellulose (NC) has also gained significant attention in pharmaceutical and biomedical applications recently, above all in wound healing (Napavichayanun et al 2016), tissue engineering (Gorgieva et al 2017;Markstedt et al 2015;Brown et al 2013), cell therapy (Lou et al 2014), gene (Ndong Ntoutoume et al 2017), drug delivery (Barbosa et al 2016;Dong et al 2014;Villanova et al 2011;Lin et al 2016), and diagnostics (Edwards et al 2016). Such a wide application spectrum is related mainly to their nanometer-sized features, large surface area, specific biomechanical characteristics, surface chemistry, ease of conjugation, high biocompatibility, and low (if any) cytotoxicity (Alexandrescu et al 2013) with tolerogenic potential to the immune system (Tomić et al 2016).…”

Supramolecular Nanofibrillar Polymer Hydrogels

“…For example, the biophysical properties of filamentous collagenous hydrogels affect interactions between cells and their extracellular matrix [13,14], while the mechanical properties of fibrin gels influence the obstruction of blood vessels with blood clots [15]. Fundamental and applied studies of nanofibrillar hydrogels are needed to further develop existing and potential novel applications of man-made nanofibrillar gels such as scaffolds in tissue engineering [16], artificial extracellular matrices for cell encapsulation [17,18], and cargo carriers in pharmaceutics and cosmetics [19][20][21][22][23]. Second, current interest in intelligent soft materials also embraces the field of nanofibrillar hydrogels.…”