Efficient Decellularization by Application of Moderate High Hydrostatic Pressure with Supercooling Pretreatment
| Autor: | Shogo Miyata, Masashi Yamamoto, Daiki Zemmyo |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Decellularization
Chemistry high hydrostatic pressure Mechanical Engineering Hydrostatic pressure cyclic hydrostatic pressure preservation of structure supercooling Article Cell membrane medicine.anatomical_structure Control and Systems Engineering Cell culture Collagen network TJ1-1570 Biophysics Extracellular medicine decellularization collagen structure Denaturation (biochemistry) Mechanical engineering and machinery Electrical and Electronic Engineering Supercooling |
| Zdroj: | Micromachines Micromachines; Volume 12; Issue 12; Pages: 1486 Micromachines, Vol 12, Iss 1486, p 1486 (2021) |
| ISSN: | 2072-666X |
| Popis: | Decellularized tissues are considered superior scaffolds for cell cultures, preserving the microstructure of native tissues and delivering many kinds of cytokines. High hydrostatic pressure (HHP) treatment could remove cells physically from biological tissues rather than chemical methods. However, there are some risks of inducing destruction or denaturation of extracellular matrices (ECMs) at an ultrahigh level of HHP. Therefore, efficient decellularization using moderate HHP is required to remove almost all cells simultaneously to suppress tissue damage. In this study, we proposed a novel decellularization method using a moderate HHP with supercooling pretreatment. To validate the decellularization method, a supercooling device was developed to incubate human dermal fibroblasts or collagen gels in a supercooled state. The cell suspension and collagen gels were subjected to 100, 150, and 200 MPa of HHP after supercooling pretreatment, respectively. After applying HHP, the viability and morphology of the cells and the collagen network structure of the gels were evaluated. The viability of cells decreased dramatically after HHP application with supercooling pretreatment, whereas the microstructures of collagen gels were preserved and cell adhesivity was retained after HHP application. In conclusion, it was revealed that supercooling pretreatment promoted the denaturation of the cell membrane to improve the efficacy of decellularization using static application of moderate HHP. Furthermore, it was demonstrated that the HHP with supercooling pretreatment did not degenerate and damage the microstructure in collagen gels. |
| Databáze: | OpenAIRE |
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