A multiple-funnels cell culture insert for the scale-up production of uniform cell spheroids
| Autor: | Masako Kawagoe, Kai Chiang Yang, Shoichiro Sumi, Goichi Yanai, Yasumasa Shirouzu, Rie Abe |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Materials science Embryoid body genetic structures Cell Biomedical Engineering Nanotechnology Cell sphere Suspension culture Insert (molecular biology) Hanging drop Biomaterials 03 medical and health sciences medicine Cell culture insert lcsh:QH573-671 Mouse embryonic stem cell lcsh:R5-920 lcsh:Cytology Spheroid equipment and supplies Uniform size 030104 developmental biology medicine.anatomical_structure Cell culture SCALE-UP SPHERES sense organs lcsh:Medicine (General) Developmental Biology |
| Zdroj: | Regenerative Therapy, Vol 7, Iss C, Pp 52-60 (2017) |
| ISSN: | 2352-3204 |
| Popis: | Introduction: Formation of cell spheres is an important procedure in biomedical research. A large number of high-quality cell spheres of uniform size and shape are required for basic studies and therapeutic applications. Conventional approaches, including the hanging drop method and suspension culture, are used for cell sphere production. However, these methods are time consuming, cell spheres cannot be harvested easily, and it is difficult to control the size and geometry of cell spheres. To resolve these problems, a novel multiple-funnel cell culture insert was designed for size controlling, easy harvesting, and scale-up production of cell spheres. Methods: The culture substrate has 680 micro-funnels with a 1-mm width top, 0.89 mm depth, and 0.5 mm square bottom. Mouse embryonic stem cells were used to test the newly developed device. The seeded embryonic stem cells settled at the downward medium surface toward the bottom opening and aggregated as embryoid bodies (EBs). For cell sphere harvest, the bottom of the culture insert was put in contact with the medium surface in another culture dish, and the medium in the device flowed down with cell spheres by hydrostatic pressure. Results: Compact cell spheres with uniform size and shape were collected easily. The diameter of the spheres could be controlled by adjusting the seeding cell density. Spontaneous neural differentiation (nestin and Tju1) and retinoic acid-induced endodermal differentiation (Pdx-1 and insulin I) were improved in the EBs produced using the new insert compared to those in EBs produced by suspension culture. Conclusions: This novel cell culture insert shall improve future studies of cell spheres and benefit clinical applications of cell therapy. |
| Databáze: | OpenAIRE |
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