Reversible Cell Layering for Heterogeneous Cell Assembly Mediated by Ionic Cross-Linking of Chitosan and a Functionalized Cell Surface Membrane

Seungmi Ryu, Hyunbum Kim, Seokyung Kang, Kwangsoo Shin, Seon Yeop Jung, Jiwoong Heo, Jin Han, Jeong Kee Yoon, Ju Ro Lee, Jinkee Hong, Kyung Hyun Ahn, Taeghwan Hyeon, Nathaniel Suk Yeon Hwang, Byung Soo Kim

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)


Current heterogeneous cell assembly techniques in coculture systems rely on irreversible cell layering or a cell separation membrane. However, the techniques possess major drawbacks of inefficiency in direct interactions of the assembled cell layers and cell separation following coculture, which hamper characterization and therapeutic applications of the cells following coculture. Here, we develop a reversible cell layering platform for assembly of heterogeneous cells that allows both active direct cell-cell interactions and facile cell separation. Anionic maleimide-chondroitin-sulfate is grafted onto the surface membrane of myogenic C2C12 cells and human mesenchymal stem cells (hMSCs) to modify the surface charge of the cells without cytotoxicity. A highly porous chitosan thin film is formed in situ interspacing between the heterogeneous cell layers via ionic cross-linking of cationic chitosan and anionic functionalized cells, forming compactly assembled double-layered cell constructs. The chitosan film enables layering of the cells, which allows active direct interactions between the cell layers, and facile delayering of the cells through simple treatment with mild shear stress. The developed platform promotes the myogenic commitment of hMSCs via direct contact with C2C12 cells, mimicking the interactions that trigger stem cell differentiation in vivo. Delivery of the myogenic committed cells to muscle-injured animal models shows evident muscle regeneration.

Original languageEnglish
Pages (from-to)5294-5305
Number of pages12
JournalChemistry of Materials
Issue number12
Publication statusPublished - 2017 Jun 27

Bibliographical note

Funding Information:
This study was supported by Grants HI14C3270 and HI15C3029 from the Korea Health Industry Development Institute (KHIDI) funded by the Ministry of Health and Welfare Republic of Korea. This study was also supported by the National Research Foundation (NRF) funded by the Ministry of Science, ICT, and Future Planning (NRF-2016R1E1A1A01943393 NRF-2012M3A9C6050102, and 2017R1A2B3005842).

Publisher Copyright:
© 2017 American Chemical Society.

All Science Journal Classification (ASJC) codes

  • Chemistry(all)
  • Chemical Engineering(all)
  • Materials Chemistry


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