Culture and differentiation of preadipocytes in two-dimensional and three-dimensional in vitro systems

Daya, Sandeep; Loughlin, Alison J. and MacQueen, Hilary A. (2007). Culture and differentiation of preadipocytes in two-dimensional and three-dimensional in vitro systems. Differentiation, 75(5) pp. 360–370.



Adipogenesis is a complex process that involves
the differentiation of preadipocytes into mature
adipocytes. We have developed two-dimensional (2D)
and three-dimensional (3D) cell culture systems for
the purpose of culturing and differentiating primary
preadipocytes in vitro. Differentiating preadipocytes
show multiple lipid droplet accumulation and comparable
protein expression patterns to mature adipocytes in
vivo. We report that in both in vitro systems terminally
differentiated adipocytes show characteristics similar to
those of mature adipocytes in vivo, assessed by the expression
of the S100a/b protein, insulin receptor and
caveolin-1, and receptors for inflammatory mediators,
namely tumor necrosis factor-a receptors I and II
(TNFRI and TNFRII) and chemokine receptor 5
(CCR5). Our results demonstrate that the S100 protein,
caveolin-1, and insulin receptor are expressed and upregulated
in differentiating and terminally differentiated
cells. In addition, the receptors for TNFa are not present
in preadipocytes but are expressed in differentiating
preadipocytes and in differentiated adipocytes. Similarly,
CCR5 was exclusively expressed in differentiating
preadipocytes and terminally differentiated adipocytes,
but not in preadipocytes. Both 2D and 3D culture models
are highly robust and reproducible and offer the potential
to study adipogenesis and cellular interactions
closely resembling and comparable to those in vivo. Our
3D collagen system offers a distinct advantage over the
2D system in that the adipocytes remain confined within
the matrix and remain intact during biochemical analysis.
Moreover, the collagen matrix allows adipocytes to
closely simulate morphological characteristics and behavior
as in vivo whilst permitting manipulation of the
microenvironment in vitro to study adipogenesis.

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