Artigo
Bovine dedifferentiated adipose tissue (DFAT) cells
Autor
Wei, Shengjuan
Du, Min
Jiang, Zhihua
Duarte, Marcio S.
Fernyhough- Culver, Melinda
Albrecht, Elke
Will, Katja
Zan, Linsen
Hausman, Gary J.
Elabd, Elham M. Youssef
Bergen, Werner G.
Basu, Urmila
Dodson, Michael V.
Institución
Resumen
Dedifferentiated fat cells (DFAT cells) are derived from lipid-containing (mature) adipocytes, which possess the ability
to symmetrically or asymmetrically proliferate, replicate, and redifferentiate/transdifferentiate. Robust cell isolation and
downstream culture methods are needed to isolate large numbers of DFAT cells from any (one) adipose depot in order
to establish population dynamics and regulation of the cells within and across laboratories. In order to establish more
consistent/repeatable methodology here we report on two different methods to establish viable DFAT cell cultures: both
traditional cell culture flasks and non-traditional (flat) cell culture plates were used for ceiling culture establishment.
Adipocytes (maternal cells of the DFAT cells) were easier to remove from flat culture plates than flasks and the flat
plates also allowed cloning rings to be utilized for cell/cell population isolation. While additional aspects of usage of
flat-bottomed cell culture plates may yet need to be optimized by definition of optimum bio-coating to enhance cell
attachment, utilization of flat plate approaches will allow more efficient study of the dedifferentiation process or the
DFAT progeny cells. To extend our preliminary observations, dedifferentiation of Wagyu intramuscular fat (IMF)-derived
mature adipocytes and redifferentiation ability of DFAT cells utilizing the aforementioned isolation protocols were
examined in traditional basal media/differentiation induction media (DMI) containing adipogenic inducement reagents.
In the absence of treatment approximately 10% isolated Wagyu IMF-mature adipocytes dedifferentiated spontaneously
and 70% DFAT cells displayed protracted adipogenesis 12 d after confluence in vitro. Lipid-free intracellular vesicles in the
cytoplasm (vesicles possessing an intact membrane but with no any observable or stainable lipid inside) were observed
during redifferentiation. One to 30% DFAT cells redifferentiated into lipid-assimilating adipocytes in the DMI media, with
distinct lipid-droplets in the cytoplasm and with no observable lipid-free vesicles inside. Moreover, a high confluence
level promoted the redifferentiation efficiency of DFAT cells. Wagyu IMF dedifferentiated DFAT cells exhibited unique
adipogenesis modes in vitro, revealing a useful cell model for studying adipogenesis and lipid metabolism.