Apr 1 2010
In a recent study initiated by Beiersdorf AG, Florian Spörl and
colleagues evaluated the use of Roche Applied Science's (Pink
Sheets: RHHBY) (SWX:RO) (SWX:ROG) xCELLigence System as a viable
system to monitor not only cholesterol extraction and repletion but also
cholesterol reorganization in human keratinocytes in real time. The
xCELLigence System offers a novel approach to non-invasive long-term
observation of membrane cholesterol dynamics in primary human
keratinocytes. This real-time cell analysis system measures alterations
in cellular impedance using microelectrodes on the surface of a 96-well
culture plate (E-Plate). Spörl et al. were able to analyze the effects
of membrane cholesterol reorganization on proliferation and
differentiation, and their findings imply a strictly controlled
mechanism for the regulation of membrane cholesterol composition in both
early and late keratinocyte differentiation.
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Cholesterol within biological membranes is organized in distinct
liquid-ordered micro-domains known as lipid rafts. As was recently
suggested, these lipid rafts play a role in cellular processes such as
early differentiation and apoptosis in human keratinocytes. To date,
monitoring the dynamics of cholesterol organization in plasma membranes
remains challenging - the techniques available are simply limited. Spörl
and colleagues are now the first to use the impedance-based xCELLigence
System to monitor non-invasively and in real time membrane
cholesterol reorganization and keratinocyte cell behavior. The authors
note that the use of this real-time cell analysis system will further
enhance understanding of how physiological processes in keratinocytes
are controlled by membrane cholesterol.
As a first step, the researchers evaluated that the real-time cell
analysis system was a viable tool to monitor normal cell growth of
primary keratinocytes. They then looked at cholesterol extraction and
repletion and found that alterations in cellular impedance could be
correlated with changes in membrane cholesterol. Not only did decreasing
and increasing cellular impedance values represent extraction and
repletion of cholesterol, Spörl et al. also described a correlation
between cellular impedance and cholesterol-dependent lateral mobility in
lipid rafts. They were then able to analyze physiological effects of the
reorganization of membrane cholesterol in more detail and saw that the
proliferative capacity of primary keratinocytes was increased upon
cholesterol depletion.
During the late stage of differentiation, keratinocytes undergo major
calcium-dependent morphological changes. According to the Beiersdorf
study, these differentiation-related effects could also be visualized
using cellular impedance measurements and the xCELLigence System. They
were able to further analyze the role of membrane cholesterol in late
keratinocyte differentiation and identified keratin 2 as a previously
unreported differentiation marker that is regulated by membrane
cholesterol organization. Lipid-raft mediated signaling might play an
additional role in this regulatory process. Remarkably, these results
were complemented by observations of differentiation-dependent
morphological changes using the xCELLigence System.
Source Beiersdorf AG