Generation of the Compression-induced Dedifferentiated Adipocytes (CiDAs) Using Hypertonic Medium
Yiwei Li,
Angelo Mao,
Bo Seo,
Xing Zhao,
Satish Gupta,
Maorong Chen,
Yu Han,
Ting-Yu Shih,
David Mooney,
Ming Guo
Affiliations
Yiwei Li
Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
Angelo Mao
John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USAWyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA
Bo Seo
John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USAWyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA
Xing Zhao
Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
Satish Gupta
Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
Maorong Chen
F. M. Kirby Neurobiology Center, Boston Children’s Hospital, Department of Neurology, Harvard Medical School, Boston, MA 02115, USA
Yu Han
Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
Ting-Yu Shih
John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USAWyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA
David Mooney
John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USAWyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138, USA
Ming Guo
Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
Current methods to obtain mesenchymal stem cells (MSCs) involve sampling, culturing, and expanding of primary MSCs from adipose, bone marrow, and umbilical cord tissues. However, the drawbacks are the limited numbers of total cells in MSC pools, and their decaying stemness during in vitro expansion. As an alternative resource, recent ceiling culture methods allow the generation of dedifferentiated fat cells (DFATs) from mature adipocytes. Nevertheless, this process of spontaneous dedifferentiation of mature adipocytes is laborious and time-consuming. This paper describes a modified protocol for in vitro dedifferentiation of adipocytes by employing an additional physical stimulation, which takes advantage of augmenting the stemness-related Wnt/β-catenin signaling. Specifically, this protocol utilizes a polyethylene glycol (PEG)-containing hypertonic medium to introduce extracellular physical stimulation to obtain higher efficiency and introduce a simpler procedure for adipocyte dedifferentiation.
