Journal article
Bovine dedifferentiated adipose tissue (DFAT) cells: DFAT cell isolation
Adipocyte, Vol.2(3), pp.148-159
07/01/2013
Handle:
https://hdl.handle.net/2376/111110
PMID: 23991361
Abstract
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.
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Details
- Title
- Bovine dedifferentiated adipose tissue (DFAT) cells
- Creators
- Shengjuan Wei - College of Animal Science and Technology; Northwest A&F University; Yangling, Shaanxi Province PR ChinaMin Du - Department of Animal Sciences; Washington State University; Pullman, WA USAZhihua Jiang - Department of Animal Sciences; Washington State University; Pullman, WA USAMarcio S Duarte - Department of Animal Sciences; Washington State University; Pullman, WA USAMelinda Fernyhough-Culver - Abitec Corporation; Columbus, OH USAElke Albrecht - Institute of Muscle Biology and Growth; Leibniz Institute for Farm Animal Biology; Dummerstorf, GermanyKatja Will - Institute of Muscle Biology and Growth; Leibniz Institute for Farm Animal Biology; Dummerstorf, GermanyLinsen Zan - College of Animal Science and Technology; Northwest A&F University; Yangling, Shaanxi Province PR ChinaGary J Hausman - Animal Science Department; University of Georgia; Athens, GA USAElham M Youssef Elabd - Biochemistry Department; National Research Centre; Cairo, EgyptWerner G Bergen - Program in Cellular and Molecular Biosciences/Department of Animal Sciences; Auburn University; Auburn, AL USAUrmila Basu - Department of Agriculture, Food and Nutritional Science; University of Alberta; Edmonton, AB CanadaMichael V Dodson - Department of Animal Sciences; Washington State University; Pullman, WA USA
- Publication Details
- Adipocyte, Vol.2(3), pp.148-159
- Academic Unit
- Animal Sciences, Department of
- Publisher
- Landes Bioscience
- Identifiers
- 99900547050701842
- Language
- English
- Resource Type
- Journal article