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Establishment of a preadipocyte cell line derived from mature adipocytes of GFP transgenic mice and formation of adipose tissue

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Abstract

We established a preadipocyte cell line from mature adipocytes obtained from subcutaneous fat tissue of green fluorescent protein (GFP) transgenic mice. The floating top layer, containing mature adipocytes, was isolated from subcutaneous fat tissue by collagenase digestion and filtration. Fluorescence-activated cell sorting and microscopic analysis revealed that the floating cell fraction comprised a highly homogeneous adipocyte population with no adipose stromal-vascular cells. Isolated mature adipocytes dedifferentiated into fibroblast-like cells and actively proliferated in ceiling culture. In vitro studies showed that the cells could redifferentiate into mature adipocytes in an identical way to 3T3-L1 preadipocytes. No changes in the differentiation pattern were observed during the propagation of our cells. They were successfully maintained and differentiated for at least 22 passages. We named these cells dedifferentiated fat (DFAT-GFP) cells. When DFAT-GFP cells were implanted subcutaneously into C57BL/6N mice, they developed highly vascularized fat pads that morphologically resembled normal subcutaneous adipose tissue and consisted of GFP-positive cells; however, implanted 3T3-L1 cells did not have such an effect on the mice. We conclude that DFAT-GFP cells provide a model that should enable us to study the mechanisms of adipocyte differentiation and adipose tissue formation in vivo and in vitro.

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Acknowledgments

We thank Atsuyoshi Ichimura and Asuka Iijima for technical assistance and discussion.

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Authors and Affiliations

  1. Division of Applied Life Sciences, Graduate School of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa City, Kanagawa, 252–8510, Japan

    Hiroyuki Nobusue, Tsuyoshi Endo & Koichiro Kano

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  1. Hiroyuki Nobusue
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  2. Tsuyoshi Endo
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  3. Koichiro Kano
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Corresponding author

Correspondence to Koichiro Kano.

Additional information

This work was supported by grants from the Japan Ministry of Education, Science, Sports, and Culture (no. 19580348) and from MEXT. HAITEKU (2007–2011).

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Nobusue, H., Endo, T. & Kano, K. Establishment of a preadipocyte cell line derived from mature adipocytes of GFP transgenic mice and formation of adipose tissue. Cell Tissue Res 332, 435–446 (2008). https://doi.org/10.1007/s00441-008-0593-9

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