한빛사 논문
In Young Choia,b,1, HoTae Lima,c,1, Alex Huynha, James Schofielda, Hyeon Jin Chod, Hosuk Leea, Peter Andersena, Joo Heon Shinj, Won Do Heoe,f, Sang-Hwan Hyuna,c, Yong Jun Kimg, Yohan Ohh,i,*, Hyesoo Kima,j,*, Gabsang Leea,j,k,*
aInstitute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA
bDepartment of Pathology, Graduate School, Kyung Hee University, School of Medicine, Seoul, Republic of Korea
cCollege of Veterinary Medicine, Chungbuk National University, Chungbuk, Republic of Korea
dLieber Institute for Brain Development, Johns Hopkins Medical Campus, Baltimore, MD, USA
eCenter for Cognition and Sociality, Institute for Basic Science, Daejeon, Republic of Korea
fDepartment of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea
gDepartment of Pathology, Department of Biomedical Science, Graduate School, Kyung Hee University, School of Medicine, Seoul, Republic of Korea
hDepartment of Biomedical Science, Graduate School of Biomedical Science and Engineering, Hanyang University, Seoul, Republic of Korea
iDepartment of Biochemistry and Molecular Biology, College of Medicine, Hanyang University, Seoul, Republic of Korea
jDepartment of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD, USA
kThe Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA
1These authors contributed equally to this work.
*Corresponding author
Abstract
Stem cell fate is largely determined by cellular signaling networks and is heavily dependent on the supplementation of exogenous recombinant proteins into culture media; however, uneven distribution and inconsistent stability of recombinant proteins are closely associated with the spontaneous differentiation of pluripotent stem cells (PSCs) and result in significant costs in large-scale manufacturing. Here, we report a novel PSC culture system via wirelessly controllable optical activation of the fibroblast growth factor (FGF) signaling pathway without the need for supplementation of recombinant FGF2 protein, a key molecule for maintaining pluripotency of PSCs. Using a fusion protein between the cytoplasmic region of the FGF receptor-1 and a light-oxygen-voltage domain, we achieved tunable, blue light-dependent activation of FGF signaling in human and porcine PSCs. Our data demonstrate that a highly controllable optical stimulation of the FGF signaling pathway is sufficient for long-term maintenance of PSCs, without the loss of differentiation potential into three germ layers. This culture system will be a cost-effective platform for a large-scale stem cell culture.
Keywords : Optogenetics; FGF2; FGF signaling; Pluripotent stem cell; Pluripotency
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