Li Yiwei, Mao Angelo S, Seo Bo Ri, Zhao Xing, Gupta Satish Kumar, Chen Maorong, Han Yu Long, Shih Ting-Yu, Mooney David J, Guo Ming
Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA.
Bio Protoc. 2021 Feb 20;11(4):e3920. doi: 10.21769/BioProtoc.3920.
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 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 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.
目前获取间充质干细胞（MSC）的方法包括从脂肪、骨髓和脐带组织中采集、培养和扩增原代MSC。然而，缺点是MSC库中的总细胞数量有限，并且它们在扩增过程中干性会衰减。作为一种替代资源，最近的顶空培养方法可以从成熟脂肪细胞中生成去分化脂肪细胞（DFAT）。然而，成熟脂肪细胞的这种自发去分化过程既费力又耗时。本文描述了一种通过采用额外的物理刺激来进行脂肪细胞去分化的改良方案，该方案利用增强与干性相关的Wnt/β-连环蛋白信号传导。具体而言，该方案利用含聚乙二醇（PEG）的高渗培养基引入细胞外物理刺激，以获得更高的效率，并为脂肪细胞去分化引入更简单的程序。
