Non-invasive stem cell tracking in hindlimb ischemia animal model using bio-orthogonal copper-free click chemistry.
Labeling of stem cells aims to distinguish transplanted cells from host cells, understand in vivo fate of transplanted cells, particularly important in stem cell therapy. Adipose-derived mesenchymal stem cells (ASCs) are considered as an emerging therapeutic option for tissue regeneration, but much remains to be understood regarding the in vivo evidence. In this study, a simple and efficient cell labeling method for labeling and tracking of stem cells was developed based on bio-orthogonal copper-free click chemistry, and it was applied in a mouse hindlimb ischemia model. The human ASCs were treated with tetra-acetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) to generate glycoprotein with unnatural azide groups on the cell surface, and the generated azide groups were fluorescently labeled by specific binding of dibenzylcyclooctyne-conjugated Cy5 (DBCO-Cy5). The safe and long-term labeling of the hASCs by this method was first investigated in vitro. Then the DBCO-Cy5-hASCs were transplanted into the hindlimb ischemia mice model, and we could monitor and track in vivo fate of the cells using optical imaging system. We could clearly observe the migration potent of the hASCs toward the ischemic lesion. This approach to design and tailor new method for labeling of stem cells may be useful to provide better understanding on the therapeutic effects of transplanted stem cells into the target diseases.
Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology, Hwarangno 14-gil 6, Seongbuk-gu, Seoul 136-791, Republic of Korea; Department of Chemical and Biomolecular Engineering, Yonsei University, 50 Yonseiro, Seodaemun-gu, Seoul, 120-749 Republic of Korea.
Si Yeon
SY
Lee
Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology, Hwarangno 14-gil 6, Seongbuk-gu, Seoul 136-791, Republic of Korea.
Sangmin
S
Lee
Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology, Hwarangno 14-gil 6, Seongbuk-gu, Seoul 136-791, Republic of Korea.
Jangwook
J
Lee
College of Pharmacy, Graduate School of Pharmaceutical Sciences, Ewha Womans University, Seoul 120-750, Republic of Korea.
Ji Young
JY
Yhee
Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology, Hwarangno 14-gil 6, Seongbuk-gu, Seoul 136-791, Republic of Korea; Departments of Chemical Engineering and Bionanotechnology, Hanyang University, Ansan, Gyeonggi-do 426-791, Republic of Korea.
Hwa In
HI
Yoon
Department of Medicine, School of Medicine, Konkuk University, Seoul, Republic of Korea.
Soon-Jung
SJ
Park
Department of Medical Lifescience, College of Medicine, The Catholic University of Korea, 222 Banpo-daero, Seocho-gu, Seoul 06591, Republic of Korea.
Heebeom
H
Koo
Department of Medicine, School of Medicine, Konkuk University, Seoul, Republic of Korea.
Sung-Hwan
SH
Moon
College of Pharmacy, Graduate School of Pharmaceutical Sciences, Ewha Womans University, Seoul 120-750, Republic of Korea.
Hyukjin
H
Lee
Departments of Chemical Engineering and Bionanotechnology, Hanyang University, Ansan, Gyeonggi-do 426-791, Republic of Korea.
Yong Woo
YW
Cho
Next-generation Pharmaceutical Research Center, Korea Institute of Toxicology, Daejeon, Republic of Korea.
Sun Woong
SW
Kang
Department of Chemical and Biomolecular Engineering, Yonsei University, 50 Yonseiro, Seodaemun-gu, Seoul, 120-749 Republic of Korea. Electronic address: leessy@yonsei.ac.kr.
Sang-Yup
SY
Lee
Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology, Hwarangno 14-gil 6, Seongbuk-gu, Seoul 136-791, Republic of Korea. Electronic address: kim@kist.re.kr.
Kwangmeyung
K
Kim
D000273:Adipose Tissue / Q000166:cytology; D000818:Animals; D001386:Azides / Q000737:chemistry; D058948:Cell Tracking / Q000379:methods*; D057930:Click Chemistry / Q000379:methods; D004195:Disease Models, Animal; D005456:Fluorescent Dyes / Q000737:chemistry; D006614:Hindlimb; D006801:Humans; D021621:Imaging, Three-Dimensional; D007511:Ischemia / Q000473:pathology / Q000628:therapy*; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice
D001386:Azides; D005456:Fluorescent Dyes
Bio-orthogonal copper-free click chemistry; Cell labeling and tracking; Hindlimb ischemia; Mesenchymal stem cell; Metabolic glycoengineering