Experimental design of a culture approach for corneal endothelial cells of New Zealand white rabbit
| Autor: | Jorge E. Valdez-García, Daniela Gomez-Elizondo, Cesar E. Calzada-Rodríguez, Judith Zavala, Italia Tatnaí Cárdenas-Rodríguez, Mariana Lopez-Martinez, María Dolores Montalvo-Parra, Isaac Alejandro Vidal-Paredes, Guiomar Farid Torres-Guerrero |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Corneal endothelium Cell biology Cellular differentiation ATPase Extracellular matrix 03 medical and health sciences Tissue culture 0302 clinical medicine Tissue engineering In vivo Cell differentiation lcsh:Social sciences (General) lcsh:Science (General) Multidisciplinary biology Chemistry Circularity Experimental design Ophthalmology 030104 developmental biology Cell culture Regenerative medicine biology.protein cardiovascular system lcsh:H1-99 Collagen 030217 neurology & neurosurgery Research Article lcsh:Q1-390 |
| Zdroj: | Heliyon, Vol 6, Iss 10, Pp e05178-(2020) Heliyon |
| ISSN: | 2405-8440 |
| Popis: | The harvesting of corneal endothelial cells (CEC) has received special attention due to its potential as a therapy for corneal blindness. The main challenges are related to the culture media formulation, cellular density at the primary isolation, and the number of passages in which CEC can retain their functional characteristics. To alternate different media formulations to harvest CEC has an impact on the cellular yield and morphology. Therefore, we analyzed four different sequences of growth factor-supplemented Stimulatory (S) and non-supplemented Quiescent (Q) media, upon passages to find the optimal S-Q culture sequence. We assessed cell yield, morphology, procollagen I production, Na+/K+-ATPase function, and the expression of ZO-1 and Na+/K+-ATPase. Our results show SQSQ and SQQQ sequences with a balance between an improved cell yield and hexagonal morphology rate. CEC cultured in the SQQQ sequence produced procollagen I, showed Na+/K+-ATPase function, and expression of ZO-1 and Na+/K+-ATPase. Our study sets a culture approach to guarantee CEC expansion, as well as functionality for their potential use in tissue engineering and in vivo analyses. Thus, the alternation of S and Q media improves CEC culture. SQQQ sequence demonstrated CEC proliferation and lower the cost implied in SQSQ sequences. We discarded the use of pituitary extract and ROCK inhibitors as essential for CEC proliferation. Tissue engineering; Cell culture; Cell differentiation; Extracellular matrix; Tissue culture; Cell biology; Regenerative medicine; Ophthalmology; Corneal endothelium; ATPase; Collagen; Circularity; Experimental design. |
| Databáze: | OpenAIRE |
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