Quantified forces between HepG2 hepatocarcinoma and WA07 pluripotent stem cells with natural biomaterials correlate with in vitro cell behavior

Abstract

In vitro cell culture or tissue models that mimic in vivo cellular response have potential in tissue engineering and regenerative medicine, and are a more economical and accurate option for drug toxicity tests than animal experimentation. The design of in vivo -like cell culture models should take into account how the cells interact with the surrounding materials and how these interactions affect the cell behavior. Cell-material interactions are furthermore important in cancer metastasis and tumor progression, so deeper understanding of them can support the development of new cancer treatments. Herein, the colloidal probe microscopy technique was used to quantify the interactions of two cell lines (human pluripotent stem cell line WA07 and human hepatocellular carcinoma cell line HepG2) with natural, xeno-free biomaterials of different chemistry, morphology, and origin. Key components of extracellular matrices –human collagens I and IV, and human recombinant laminin-521−, as well as wood-derived, cellulose nanofibrils –with evidenced potential for 3D cell culture and tissue engineering– were analysed. Both strength of adhesion and force curve profiles depended on biomaterial nature and cell characteristics. The successful growth of the cells on a particular biomaterial required cell-biomaterial adhesion energies above 0.23 nJ/m. The information obtained in this work supports the development of new materials or hybrid scaffolds with tuned cell adhesion properties for tissue engineering, and provides a better understanding of the interactions of normal and cancerous cells with biomaterials in the human body.

Differentiation of Human Pluripotent Stem Cells Into Definitive Endoderm Cells in Various Flexible Three-Dimensional Cell Culture Systems: Possibilities and Limitations

“…Based on the results from the 2D DE differentiation experiments showing that 6-day DE induction is the most effective method ( Bogacheva et al, 2018 ), we performed 6-day DE differentiation of iPS(IMR90)-4 cells in three 3D conditions. Suspension culture in this study represents a biomaterial/scaffold-free 3D culture condition, while NFC hydrogel is an inert biomaterial that exhibits weak nonspecific interactions with cells ( Harjumaki et al, 2019 ). Conversely, BME interacts with stem cells and DE cells directly via cell membrane receptors, so it represents an active cell-interacting biomaterial.…”

“…Decreasing E-cadherin expression during DE differentiation ( D’Amour et al, 2005 ) further reduces cell–cell interactions, thereby contributing to disrupted spheroid morphology. In contrast, NFC hydrogel has negligible adhesion forces to stem cells ( Harjumaki et al, 2019 )—minimizing cell–matrix interactions. Thus, the predominance of cell–cell interactions may account for the well maintenance of the spheroid shape in NFC hydrogel ( Figure 9 ).…”

“…Intact spheroids formed in NFC hydrogel can be harvested after removal of the hydrogel by utilizing a cellulase enzyme ( Lou et al, 2014 ), thereby facilitating various downstream analyses and applications ( Lou et al, 2014 ). NFC hydrogel displayed weak interactions with human ESCs and HepG2 cells compared with natural extracellular matrix proteins including collagens and laminins ( Harjumaki et al, 2019 ). In contrast, suspension culture is a biomaterial-free system, usually performed in a low-adhesive culture dish or a flask for scaled-up production.…”

An Efficient and Inexpensive PEI-mediated Transfection for Transient Overexpression of CYP2C9 in WRL 68 Normal Liver Cells

“…Previous study reported that variability in PEI toxicity seen in different cell lines is possibly due to the difference in physiology of the cells [18]. WRL 68 and HepG2 are both derived from liver exhibiting morphologic structures similar to hepatocytes [19,20]. Both liver derived cell lines are more affected by PEI toxicity possibly due to their comparable physiology.…”

Influence of surface topography on PCL electrospun scaffolds for liver tissue engineering

“…4 This is in part due to it's virus-free nature, high proliferation rate and it's ability to perform many hepatic specific functions such as albumin secretion. [93][94][95] However, because it is relatively stable in vitro culture, it does not give a true reflection of primary culture. Future work is needed to access primary or stem cells derived hepatocytes to verify the effect of scaffolds on long-term in vitro culture while ensuring that variables such as fibre base size and pore size are taken into count.…”