Use of nitrocellulose membranes as a scaffold in cell culture

Abstract

Nitrocellulose membranes, one of the most important and oldest cellulose derivatives, are commonly used for nucleic acid and protein detection in research and diagnostic applications. However, a limited number of studies have explored whether they can act as scaffolds for cell growth. In this study, we investigated this polymeric material for its ability to support the growth of human cells. Eight established cell lines were examined for adherence, growth, spread, and survival on nitrocellulose membranes by optical microscopy after hematoxylin and eosin and/or immunocytochemical staining and by scanning electron microscopy. Apoptosis and leakage of lactate dehydrogenase (LDH) were also assessed. All cells readily adhered to and spread on the surface of nitrocellulose membranes as well as coverslips, and the cells maintained the expression of digestive system-specific genes. No significant change was detected in apoptosis or leakage of LDH from cells grown on nitrocellulose membranes. These results suggested that nitrocellulose membranes have a suitable cytocompatibility towards human cells and that they might be used for tissueengineering scaffolds. Moreover, we demonstrate an additional and underused property of nitrocellulose of specific relevance to microscopic imaging, as it can be rendered virtually transparent, thus the cells growing on such membranes can be observed directly under an optical microscope after staining.

Construction and characterization of a multilayered gingival keratinocyte culture model: the TURK-U model

“…In the present model, we used a semi-permeable nitrocellulose membrane as a scaffold. Nitrocellulose membranes are cellulose derivatives and their high cytocompatibility towards human cells has been shown (Li et al 2013). In our model, the semipermeable character of nitrocellulose allows the growth medium to diffuse through the membrane and to feed the epithelial cells while floating on the semi-solid medium and keeping the cells in contact to air.…”

“…In our model, the semipermeable character of nitrocellulose allows the growth medium to diffuse through the membrane and to feed the epithelial cells while floating on the semi-solid medium and keeping the cells in contact to air. Moreover, the three-dimensional structure of the porous nitrocellulose membrane provides a greater binding capacity to epithelial cells (Li et al 2013). Several tests were performed during the development process of the present model.…”

Low-cost assays in paper-based microfluidic biomedical devices

“…Another important type of paper substrate is the hydrophobic nitrocellulose membranes, which are smooth and with reasonably uniform pore size (0.45 μm) that allows for a more stable and reproducible liquid fl ow. They have been used for a long time for antigen immobilization (Hawkes et al , 1982) in, for instance, dot-immunobinding assays , because they manifest a high degree of nonspecifi c binding towards biomolecules, making them suitable for immobilization of enzymes (Martinez et al , 2008a;Lu et al , 2009aLu et al , , 2009b, proteins (Fenton et al , 2008), DNA (Cretich et al , 2010), and cells (Li et al , 2013). Besides, pores in nitrocellulose can perform purifi cation based on size by fi ltering impurities of larger diameter than the analyte, which can mitigate interferences during the assay (Abe et al , 2010).…”

“…Microfl uidic paper devices have the potential to be widely adopted analytical tools with attractive features such as low cost, portability, disposability, limited waste, simplicity, speed, lower consumption of reagents and sample, and ease of fabrication and use (Auroux et al , 2002;Reyes et al , 2002;Coltro et al , 2010;Li et al , 2013;Liana et al , 2012). Their downsized reaction zones compared to dipstick tests make them attractive when working with scarce or hard-to-get samples (Abe et al , 2008).…”

“…In fact, at least half of it is retained within the paper matrix or evaporates in the air before reaching the detection zones, resulting in less concentrated, thus harder to detect, analytes (Li et al , 2013). Losses are particularly undesirable when quantities of samples are limited.…”