In-Plate Cryopreservation of 2D and 3D Cell Models: Innovative Tools for Biomedical Research and Preclinical Drug Discovery
| Autor: | Marie-Ann Ewart, Colin J. Wilde, Alessandra Prinelli, Catarina Silva-Almeida, Erin Sutherland, Anna Pasotti, Aikaterini Telopoulou, Sophie Dunlop, Elfi Töpfer, Martin Lynch, Sisely Parks |
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| Přispěvatelé: | Prinelli A., Silva-Almeida C., Parks S., Pasotti A., Telopoulou A., Dunlop S., Sutherland E., Lynch M., Ewart M.-A., Wilde C.J., Topfer E. |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Cell physiology 3D culture Biomedical Research Computer science organoid Cell cell-based assay Cell Culture Techniques Drug Evaluation Preclinical Computational biology Biochemistry Cryopreservation Analytical Chemistry 03 medical and health sciences 0302 clinical medicine Drug Discovery medicine Humans Viability assay Primary cell multiwell platform Drug discovery Mesenchymal stem cell 030104 developmental biology medicine.anatomical_structure Cell culture in-plate cryopreservation Molecular Medicine 030217 neurology & neurosurgery Biotechnology |
| Zdroj: | SLAS discovery : advancing life sciences RD. 26(1) |
| ISSN: | 2472-5560 |
| Popis: | Cell-based assays performed in multiwell plates are utilized in basic and translational research in a variety of cell models. The assembly of these multiwell platforms and their use is often laboratory specific, preventing the standardization of methods and the comparison of outputs across different analytical sites. Moreover, when cell models are based on primary cells with specialized culture requirements, including three-dimensional (3D) cell culture, their complexity and the need for manipulation by experienced operators can add significant cost and introduce long lead times to analysis, both of which are undesirable in any preclinical situation. To address this issue, we explored adaptations of cryopreservation technology that allow cells to be cryopreserved in-plate, ready for use in analysis, and have developed a method applicable to cells from different origins and different culture formats. Here we describe the application of this technology to conventional two-dimensional (2D) monolayers of human mesenchymal stem cells (MSCs) and human macrophages derived from primary monocytes, and to 3D cultures of hepatic organoids, colon organoids, and colon tumor organoids, each presented for cryopreservation in their obligate extracellular matrix. We demonstrated that cell viability, cell physiology, and cytotoxic sensitivity were maintained after cryopreservation, such that the models offer the means to uncouple model assembly from analytical use and to standardize cell models in product form for distribution to end users. |
| Databáze: | OpenAIRE |
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