Spectroscopic label-free microscopy of changes in live cell chromatin and biochemical composition in transplantable organoids
| Autor: | Yuri Zakharov, Lev T. Perelman, Giuseppe Pettinato, Paul Kumar Upputuri, Robert A. Fisher, Umar Khan, Irving Itzkan, Liming Chen, Lei Zhang, Conor J. Sheil, Maria Glyavina, Mark F. Coughlan, Edward Vitkin, Le Qiu, Xuejun Zhang, Antonino B. D'Assoro |
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| Rok vydání: | 2021 |
| Předmět: |
Confocal
Cell Induced Pluripotent Stem Cells Microscopy medicine Biochemical composition Organoid Humans Research Articles Label free Multidisciplinary Chemistry fungi food and beverages Endothelial Cells SciAdv r-articles Cell Differentiation Optics Chromatin Cell biology Organoids medicine.anatomical_structure Homogeneous Research Article Developmental Biology |
| Zdroj: | Science Advances |
| ISSN: | 2375-2548 |
| Popis: | Formation of transplantable reproducible hiPSC organoids can be aided with real-time label-free intracellular spectroscopy. Organoids formed from human induced pluripotent stem cells (hiPSCs) could be a limitless source of functional tissue for transplantations in many organs. Unfortunately, fine-tuning differentiation protocols to form large quantities of hiPSC organoids in a controlled, scalable, and reproducible manner is quite difficult and often takes a very long time. Recently, we introduced a new approach of rapid organoid formation from dissociated hiPSCs and endothelial cells using microfabricated cell-repellent microwell arrays. This approach, when combined with real-time label-free Raman spectroscopy of biochemical composition changes and confocal light scattering spectroscopic microscopy of chromatin transition, allows for monitoring live differentiating organoids without the need to sacrifice a sample, substantially shortening the time of protocol fine-tuning. We used this approach to both culture and monitor homogeneous liver organoids that have the main functional features of the human liver and which could be used for cell transplantation liver therapy in humans. |
| Databáze: | OpenAIRE |
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