Abstract
Proteinases that digest the extracellular matrix are usually used to harvest cells from culture vessels in a general culture process, which lowers the initial adhesion rate in regenerative medicine. Cell sheet engineering is one of the most important technologies in this field, especially for transplantation, because fabricated cell sheets have rich extracellular matrixes providing strong initial adhesion. Current cell sheet fabrication relies on temperature-responsive polymer-coated dishes. Cells are cultured on such specialized dishes and subjected to low temperature. Thus, we developed a simple but versatile cell sheet fabrication method using ubiquitous culture dishes/flasks without any coating or temperature modulation. Confluent mouse myoblasts (C2C12 cell line) were exposed to ultrasonic vibration from underneath and detached as cell sheets from entire culture surfaces. Because of the absence of low temperature, cell metabolism was statically increased compared with the conventional method. Furthermore, viability, morphology, protein expression, and mRNA expression were normal. These analyses indicated no side effects of ultrasonic vibration exposure. Therefore, this novel method may become the standard for cell sheet fabrication. Our method can be easily conducted following a general culture procedure with a typical dish/flask, making cell sheets more accessible to medical experts.
Highlights
Proteinases that digest the extracellular matrix are usually used to harvest cells from culture vessels in a general culture process, which lowers the initial adhesion rate in regenerative medicine
We demonstrated that a cell sheet of mouse myoblasts could be detached from clinically ubiquitous cell culture by suitable ultrasonic exposure, which are commonly used in proof-of-concept tissue engineering and cell sheet studies[9]
The metabolism of the cell sheet fabricated by the developed method was statically improved compared with the conventional method, while cell viability, protein expression, and mRNA expression showed no significant differences
Summary
Proteinases that digest the extracellular matrix are usually used to harvest cells from culture vessels in a general culture process, which lowers the initial adhesion rate in regenerative medicine. In addition to using temperature-responsive culture dishes, ECM-rich cell sheets have been fabricated by other techniques such as using electro- and photo-responsive materials, modulation of medium, and infusing polymers[8,17,18,19,20]. These alternatives for detaching a cell sheet may overcome the two major problems of pNIPAAm-coated dishes. They still require specialized processing of a cell culture surface or chemicals that could be harmful to cells. A simple cell sheet fabrication method capable of generating an active and ECM-rich cell sheet is highly desired for the advancement of regenerative medicine
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
