Crucial factor causing collapse and aggregation of cultured cells in epon resin.

Ultrastructural artifacts regarding collapse and aggregation of cultured cells have been problematic, especially when investigated apoptotic cells. The infiltration process during sample preparation is considered to be the most crucial factor for this problem. This study was conducted using two culture systems: a suspension culture system of human T-lymphocyte Jurkat cells and rabbit mature dendritic cells and a monolayer culture system of human lung macrophages, human breast cancer cells (A-546 cells) and cat bone-invasive gingival cancer cells (sccf3 cells). Fixation was conducted prior to removing or detaching the cells from the culture dishes. Initial infiltration with a 1 : 3 volume ratio of epon resin : propylene oxide was found to be the most crucial step among these cultured cells. The improved epon-resin infiltration method could eliminate the artifacts. Thus, differentiation between artifactual images and true images is highly possible.

Efficacy of self-assembled hydrogels composed of positively or negatively charged peptides as scaffolds for cell culture.

KASEA16(+) and KASEA16(-) peptides, the net charges of which are positive and negative, respectively, under a neutral condition could undergo self-assembly into nanofibers and form transparent hydrogels without peptide aggregation upon rapid pH neutralization. The numbers of NIH3T3 cells attached to the KASEA16(+) hydrogel and KASEA16(-) hydrogel were similar, and cells proliferated with time on both hydrogels. Cells on the KASEA16(+) hydrogel had spindle-like morphology, while cells on the KASEA16(-) hydrogel formed clusters without extending cytoplasmic processes. Comparison of differently charged peptides under a neutral condition suggested that the charges of the scaffolds should be taken into consideration for the best design and selection of scaffolds for cell culture. Since the KASEA16(+) peptide could form a stable hydrogel under a neutral condition and the hydrogel served as a scaffold for cell proliferation, the KASEA16(+) hydrogel will be a useful scaffold for cell culture.