Intravital imaging and single cell transcriptomic analysis for engraftment of mesenchymal stem cells in an animal model of interstitial cystitis/bladder pain syndrome

• Published in: Biomaterials 2022-01, Vol.280, p.121277-121277, Article 121277
• Main Authors: Yu, Hwan Yeul, Lee, Seungun, Ju, Hyein, Kim, Youngkyu, Shin, Jung-Hyun, Yun, HongDuck, Ryu, Chae-Min, Heo, Jinbeom, Lim, Jisun, Song, Sujin, Lee, Sanghwa, Hong, Ki-Sung, Chung, Hyung-Min, Kim, Jun Ki, Choo, Myung-Soo, Shin, Dong-Myung
• Format: Article
• Language: English
• Subjects: Animals; Cystitis, Interstitial - therapy; Disease Models, Animal; FOS; Interstitial cystitis/bladder pain syndrome; Intravital imaging; Intravital Microscopy; Mesenchymal stem cell; Mesenchymal Stem Cell Transplantation - methods; Mesenchymal Stem Cells; Single cell analysis; Transcriptome
• ISSN: 0142-9612; 1878-5905
• DOI: 10.1016/j.biomaterials.2021.121277

Mesenchymal stem cell (MSC) therapy is a promising treatment for various intractable disorders including interstitial cystitis/bladder pain syndrome (IC/BPS). However, an analysis of fundamental characteristics driving in vivo behaviors of transplanted cells has not been performed, causing debates about rational use and efficacy of MSC therapy. Here, we implemented two-photon intravital imaging and single cell transcriptome analysis to evaluate the in vivo behaviors of engrafted multipotent MSCs (M-MSCs) derived from human embryonic stem cells (hESCs) in an acute IC/BPS animal model. Two-photon imaging analysis was performed to visualize the dynamic association between engrafted M-MSCs and bladder vasculature within live animals until 28 days after transplantation, demonstrating the progressive integration of transplanted M-MSCs into a perivascular-like structure. Single cell transcriptome analysis was performed in highly purified engrafted cells after a dual MACS−FACS sorting procedure and revealed expression changes in various pathways relating to pericyte cell adhesion and cellular stress. Particularly, FOS and cyclin dependent kinase-1 (CDK1) played a key role in modulating the migration, engraftment, and anti-inflammatory functions of M-MSCs, which determined their in vivo therapeutic potency. Collectively, this approach provides an overview of engrafted M-MSC behavior in vivo, which will advance our understanding of MSC therapeutic applications, efficacy, and safety. Using two-photon intravital imaging (IVM) and single cell transcriptome analysis, this study analyzed the in vivo behavior of engrafted MSCs and their dynamic interplay with the vasculature in injured bladders at the single-cell level. Two-photon IVM longitudinally monitored engrafted MSC in the bladder of live animals for 28 days after transplantation. The engrafted MSCs (green) were found to interact with the bladder vasculature (red) and progressively integrate as perivascular cells. Single cell analysis of the engrafted MSCs showed their pericyte nature. Among the genes characterizing the engrafted MSCs, FOS and CDK1 significantly modulated the in vitro core functions of MSCs and determined the in vivo engraftment and therapeutic potency of MSCs in the injured bladder. MSC; Mesenchymal stem cells, FOS; Fos Proto-Oncogene, CDK1; cyclin dependent kinase-1. [Display omitted]