Changes in nuclear shape and mitochondrial structure do not accompany the loss of division potential in human fibroblasts in vitro.

Abstract

Previous studies on ultrastructural changes that occur in cultured human fibroblasts during their in vitro life-span indicate that "senescent" cells characteristically possess structurally altered mitochondria, highly lobed nuclei, and an abundance of secondary lysosomes when compared to early passage cells. In the present study, we demonstrate that improper preparative methods can induce altered mitochondrial morphology in preparations of both IMR-90 and HF730A fibroblasts, regardless of passage level. We also show that nuclei of both living and fixed IMR-90 fibroblasts are ovoid in shape, not lobulate, in well-spread cells, regardless of either the passage level or the proliferative capacity of the cell. Fibroblasts contain lobulated nuclei only when they have not spread completely on the culture substrate. Lobulations can be induced at any passage level by collagenase/trypsin or trypsin/EDTA treatment prior to fixation, but not by cytochalasin B treatment or by cold temperatures. We conclude that any treatment that affects cytoskeleton-membrane-culture substrate interactions will induce this aberrant nuclear morphology, but that this is not indicative of "senescence" and does not relate to proliferative decline.