Improved methods for the differentiation of hypothalamic vasopressin neurons using mouse induced pluripotent stem cells

• Published in: Stem cell research Vol. 40; p. 101572
• Main Authors: Mitsumoto, Kazuki, Suga, Hidetaka, Sakakibara, Mayu, Soen, Mika, Yamada, Tomiko, Ozaki, Hajime, Nagai, Takashi, Kano, Mayuko, Kasai, Takatoshi, Ozone, Chikafumi, Ogawa, Koichiro, Sugiyama, Mariko, Onoue, Takeshi, Tsunekawa, Taku, Takagi, Hiroshi, Hagiwara, Daisuke, Ito, Yoshihiro, Iwama, Shintaro, Goto, Motomitsu, Banno, Ryoichi, Arima, Hiroshi
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 01.10.2019; Elsevier
• Subjects: Animals; AVP neuron; Cell Differentiation; Cell Line - cytology; Cell Line - metabolism; Cells, Cultured; Differentiation; Fibroblast Growth Factor 8 - metabolism; Fibroblasts - cytology; Fibroblasts - metabolism; Hypothalamus; Hypothalamus - cytology; Hypothalamus - metabolism; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; Maintenance; Mice; Mice, Inbred C57BL; Mouse iPS cells; Neurons - cytology; Neurons - metabolism; Vasopressins - metabolism; Mouse iPS cells; AVP neuron; Hypothalamus; Maintenance; Differentiation
• ISSN: 1873-5061; 1876-7753
• DOI: 10.1016/j.scr.2019.101572

High differentiation efficiency is one of the most important factors in developing an in vitro model from pluripotent stem cells. In this report, we improved the handling technique applied to mouse-induced pluripotent stem (iPS) cells, resulting in better differentiation into hypothalamic vasopressin (AVP) neurons. We modified the culture procedure to make the maintenance of iPS cells in an undifferentiated state much easier. Three-dimensional floating culture was demonstrated to be effective for mouse iPS cells. We also improved the differentiation method with regards to embryology, resulting in a greater number of bigger colonies of AVP neurons differentiating from mouse iPS cells. Fgf8, which was not used in the original differentiation method, increased iPS differentiation into AVP neurons. These refinements will be useful as a valuable tool for the modeling of degenerative disease in AVP neurons in vitro using disease-specific iPS cells in future studies. •A modified maintenance procedure using floating cultures of iPS cells was developed.•The spheres of iPS cells could be stored frozen.•Fgf8 treatment resulted in high differentiation efficiency into AVP neurons.