Development of non-adherent cell-enclosing domes with enzymatically cross-linked hydrogel shell
Non-adherent cells, such as hematopoietic cells and lymphocytes, are important research subjects in medical and biological fields. Therefore, a system that enables the handling of non-adherent cells in solutions in the same manner as that of adhering cells during medium exchange, exposure to chemica...
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| Published in | Biofabrication Vol. 15; no. 1; pp. 15002 - 15011 |
|---|---|
| Main Authors | , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
England
IOP Publishing
01.01.2023
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| Subjects | |
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| Abstract | Non-adherent cells, such as hematopoietic cells and lymphocytes, are important research subjects in medical and biological fields. Therefore, a system that enables the handling of non-adherent cells in solutions in the same manner as that of adhering cells during medium exchange, exposure to chemicals, washing, and staining in imaging applications would be useful. Here, we report a ‘Cell Dome’ platform in which non-adherent cells can be enclosed and grown in the cavities of about 1 mm diameter and 270
μ
m height. The domes consist of an alginate-based hydrogel shell of 90
μ
m thickness. Cell Domes were formed on glass plates by horseradish peroxidase-mediated cross-linking. Human leukaemia cell line K562 cells enclosed in Cell Domes were stable for 29 days with every 2–3 days of medium change. The enclosed cells grew in the cavities and were stained and differentiated with reagents supplied from the surrounding medium. Additionally, K562 cells that filled the cavities (a 3D microenvironment) were more hypoxic and highly resistant to mitomycin C than those cultured in 2D. These findings demonstrate that the ‘Cell Dome’ may be a promising tool for conveniently culturing and evaluating non-adherent cells. |
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| AbstractList | Non-adherent cells, such as hematopoietic cells and lymphocytes, are important research subjects in medical and biological fields. Therefore, a system that enables the handling of non-adherent cells in solutions in the same manner as that of adhering cells during medium exchange, exposure to chemicals, washing, and staining in imaging applications would be useful. Here, we report a 'Cell Dome' platform in which non-adherent cells can be enclosed and grown in the cavities of about 1 mm diameter and 270
m height. The domes consist of an alginate-based hydrogel shell of 90
m thickness. Cell Domes were formed on glass plates by horseradish peroxidase-mediated cross-linking. Human leukaemia cell line K562 cells enclosed in Cell Domes were stable for 29 days with every 2-3 days of medium change. The enclosed cells grew in the cavities and were stained and differentiated with reagents supplied from the surrounding medium. Additionally, K562 cells that filled the cavities (a 3D microenvironment) were more hypoxic and highly resistant to mitomycin C than those cultured in 2D. These findings demonstrate that the 'Cell Dome' may be a promising tool for conveniently culturing and evaluating non-adherent cells. Non-adherent cells, such as hematopoietic cells and lymphocytes, are important research subjects in medical and biological fields. Therefore, a system that enables the handling of non-adherent cells in solutions in the same manner as that of adhering cells during medium exchange, exposure to chemicals, washing, and staining in imaging applications would be useful. Here, we report a ‘Cell Dome’ platform in which non-adherent cells can be enclosed and grown in the cavities of about 1 mm diameter and 270 μ m height. The domes consist of an alginate-based hydrogel shell of 90 μ m thickness. Cell Domes were formed on glass plates by horseradish peroxidase-mediated cross-linking. Human leukaemia cell line K562 cells enclosed in Cell Domes were stable for 29 days with every 2–3 days of medium change. The enclosed cells grew in the cavities and were stained and differentiated with reagents supplied from the surrounding medium. Additionally, K562 cells that filled the cavities (a 3D microenvironment) were more hypoxic and highly resistant to mitomycin C than those cultured in 2D. These findings demonstrate that the ‘Cell Dome’ may be a promising tool for conveniently culturing and evaluating non-adherent cells. Non-adherent cells, such as hematopoietic cells and lymphocytes, are important research subjects in medical and biological fields. Therefore, a system that enables the handling of non-adherent cells in solutions in the same manner as that of adhering cells during medium exchange, exposure to chemicals, washing, and staining in imaging applications would be useful. Here, we report a 'Cell Dome' platform in which non-adherent cells can be enclosed and grown in the cavities of about 1 mm diameter and 270μm height. The domes consist of an alginate-based hydrogel shell of 90μm thickness. Cell Domes were formed on glass plates by horseradish peroxidase-mediated cross-linking. Human leukaemia cell line K562 cells enclosed in Cell Domes were stable for 29 days with every 2-3 days of medium change. The enclosed cells grew in the cavities and were stained and differentiated with reagents supplied from the surrounding medium. Additionally, K562 cells that filled the cavities (a 3D microenvironment) were more hypoxic and highly resistant to mitomycin C than those cultured in 2D. These findings demonstrate that the 'Cell Dome' may be a promising tool for conveniently culturing and evaluating non-adherent cells.Non-adherent cells, such as hematopoietic cells and lymphocytes, are important research subjects in medical and biological fields. Therefore, a system that enables the handling of non-adherent cells in solutions in the same manner as that of adhering cells during medium exchange, exposure to chemicals, washing, and staining in imaging applications would be useful. Here, we report a 'Cell Dome' platform in which non-adherent cells can be enclosed and grown in the cavities of about 1 mm diameter and 270μm height. The domes consist of an alginate-based hydrogel shell of 90μm thickness. Cell Domes were formed on glass plates by horseradish peroxidase-mediated cross-linking. Human leukaemia cell line K562 cells enclosed in Cell Domes were stable for 29 days with every 2-3 days of medium change. The enclosed cells grew in the cavities and were stained and differentiated with reagents supplied from the surrounding medium. Additionally, K562 cells that filled the cavities (a 3D microenvironment) were more hypoxic and highly resistant to mitomycin C than those cultured in 2D. These findings demonstrate that the 'Cell Dome' may be a promising tool for conveniently culturing and evaluating non-adherent cells. |
| Author | Nakahata, Masaki Qu, Yanfei Kojima, Masaru Fujita, Satoshi Sato, Ryuta Kazama, Ryotaro Fujiwara, Hiroyuki Sakai, Shinji |
| Author_xml | – sequence: 1 givenname: Ryotaro surname: Kazama fullname: Kazama, Ryotaro organization: Graduate School of Engineering Science, Osaka University , 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan – sequence: 2 givenname: Ryuta surname: Sato fullname: Sato, Ryuta organization: Graduate School of Engineering Science, Osaka University , 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan – sequence: 3 givenname: Hiroyuki surname: Fujiwara fullname: Fujiwara, Hiroyuki organization: Graduate School of Engineering Science, Osaka University , 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan – sequence: 4 givenname: Yanfei surname: Qu fullname: Qu, Yanfei organization: Graduate School of Engineering Science, Osaka University , 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan – sequence: 5 givenname: Masaki surname: Nakahata fullname: Nakahata, Masaki organization: Graduate School of Science, Osaka University , 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, Japan – sequence: 6 givenname: Masaru surname: Kojima fullname: Kojima, Masaru organization: Graduate School of Engineering Science, Osaka University , 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan – sequence: 7 givenname: Satoshi surname: Fujita fullname: Fujita, Satoshi organization: AIST-Osaka University Advanced Photonics and Biosensing Open Innovation Laboratory, National Institute of Advanced Industrial Science and Technology (AIST) , Suita, Osaka 565-0871, Japan – sequence: 8 givenname: Shinji orcidid: 0000-0002-1041-4798 surname: Sakai fullname: Sakai, Shinji organization: Graduate School of Engineering Science, Osaka University , 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan |
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| Keywords | alginate K562 cell microcapsule microdome Cell Dome non-adherent cell |
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| SubjectTerms | alginate Alginates - metabolism Cell Dome Horseradish Peroxidase Humans Hydrogels K562 cell microcapsule microdome Mitomycin non-adherent cell |
| Title | Development of non-adherent cell-enclosing domes with enzymatically cross-linked hydrogel shell |
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