Novel Egg White–based 3-D Cell Culture System

Abstract

Although three dimensional (3-D) cell culture systems have numerous advantages over traditional monolayer culture, the currently available 3-D cell culture media are cost-prohibitive for regular use by the majority of research laboratories. Here we show a simple system based on avian egg white that supports growth of cells in 3-D, at a significantly decreased cost. Specifically, we show that growth of immortalized human breast epithelial cells (MCF10A) in egg white-based medium results in formation of acini with hollow lumens, apoptotic clearance of the cells in the lumen, and apicobasal polarization comparable to what has been described using established 3-D culture media such as reconstituted basement membrane preparations (BM). There was no significant difference in MCF10A proliferation and acinar size between egg white and BM. We also cultured different established cell lines, oncogene-transformed MCF10A, and mouse mammary epithelial cells in egg white and BM, and observed similar morphology. In summary, our data convincingly argue that egg white can be used as a suitable alternative model for 3-D cell culture studies. We strongly believe that this simple and inexpensive method should allow researchers to perform 3-D cell culture experiments on a regular basis, and result in a dramatic increase of use of the 3-D cell culture in research. Thus, this finding lays the foundation for significantly increased, cost-effective use of 3-D cultures in cell biology.

In vitro 3D angiogenesis assay in egg white matrix: comparison to Matrigel, compatibility to various species, and suitability for drug testing

“…According to the previously reported method, 15 egg outer surface was thoroughly washed with ethanol 70%. Under a laminar flow hood, a small hole in the shell was made using sterilized forceps then the egg white was poured into a becher.…”

“…Consequently, it was more difficult to make the egg white become semi-solid after heating to render it a proper matrix for cell culture. We followed a previously described procedure 15 to extract the white from the eggs. By pooling the white of a large number of eggs simultaneously, we obtained enough egg white to carry out experiments over a long period of time with a homogenous cell culture matrix.…”

“…Heating time has to be adapted depending on the egg white volume seeded into the wells between 20 and 45 min for 8-well chamber slides to 12-well plates, respectively. The use of a hybridization oven seemed to produce more homogeny in the egg white heating compared with the heating block previously described, 15 enabling the use of egg white matrix in various cell culture devices.…”

“…15 In order to develop a cost-effective and multipurpose angiogenesis assay, we checked whether egg white could constitute a proper substitute of ECM for vascular cell culture. We used cells from human and rat origin to test their growth and migration abilities in such a matrix.…”

Selective comparison of gelling agents as neural cell culture matrices for long-term microelectrode array electrophysiology

“…There are several categories of gelling agents to choose from. Among those are purified peptides or (glyco)proteins of animal origin (e.g., egg white (Kaipparettu et al, 2008), proteoglycans, fibronectin, laminins, collagens or a mix of basal lamina proteins (Yurchenco, 2011)), artificial protein mixes (Zhao and Zhang, 2006), microbial-(e.g., gellan gum, xanthan gum), algae-(e.g., carrageenans, alginates, agarose) or plant-derived polymers (e.g., guar gum, locust bean gum, tragacanth) or polymers of completely synthetic origin (Kopecek, 2002;Lei and Schaffer, 2013). Although synthetic gels are usually expensive, they can be produced in a defined stoichiometry without impurities and their physico-chemical properties be tailored to the cells' needs, thereby giving more reproducible results.…”