دورية أكاديمية
Improving Endothelial Explant Tissue Culture by Novel Thermoresponsive Cell Culture System.
| العنوان: | Improving Endothelial Explant Tissue Culture by Novel Thermoresponsive Cell Culture System. |
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| المؤلفون: | Miron A; Netherlands Institute for Innovative Ocular Surgery , Rotterdam, The Netherlands.; Melles Cornea Clinic , Rotterdam, The Netherlands., Spinozzi D; Netherlands Institute for Innovative Ocular Surgery , Rotterdam, The Netherlands., Lie J; Netherlands Institute for Innovative Ocular Surgery , Rotterdam, The Netherlands.; Amnitrans EyeBank , Rotterdam, The Netherlands., Melles GR; Netherlands Institute for Innovative Ocular Surgery , Rotterdam, The Netherlands.; Melles Cornea Clinic , Rotterdam, The Netherlands.; Amnitrans EyeBank , Rotterdam, The Netherlands., Oellerich S; Netherlands Institute for Innovative Ocular Surgery , Rotterdam, The Netherlands., Ni Dhubhghaill S; Netherlands Institute for Innovative Ocular Surgery , Rotterdam, The Netherlands.; Antwerp University Hospital , Antwerp, Belgium. |
| المصدر: | Current eye research [Curr Eye Res] 2021 Mar; Vol. 46 (3), pp. 290-293. Date of Electronic Publication: 2020 Jul 29. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Informa Healthcare Country of Publication: England NLM ID: 8104312 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1460-2202 (Electronic) Linking ISSN: 02713683 NLM ISO Abbreviation: Curr Eye Res Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: London : Informa Healthcare Original Publication: London : IRL Press, [c1981- |
| مواضيع طبية MeSH: | Tissue Donors* , Visual Acuity*, Cell Culture Techniques/*methods , Descemet Stripping Endothelial Keratoplasty/*methods , Endothelium, Corneal/*cytology , Fuchs' Endothelial Dystrophy/*surgery, Cell Movement ; Cell Survival ; Cells, Cultured ; Fuchs' Endothelial Dystrophy/diagnosis ; Humans |
| مستخلص: | Aim: Studying cell migration of corneal endothelial cells in vitro is challenging because the capacity for cell migration needs to be maintained while at the same time the tissue must remain fixed on a rigid substrate. In this study, we report a thermoresponsive culture technique designed to maintain cellular viability, and to reduce tissue handling in order to analyze in vitro endothelial cell migration from corneal grafts. Materials and Methods: As a test tissue, fifteen Quarter-Descemet membrane endothelial keratoplasty (Q-DMEK) grafts were used that were embedded in a three-dimensional culture system using a temperature-reversible hydrogel and cultured over 2-3 weeks in a humidified atmosphere at 37°C and 5% CO Results: All grafts could be successfully cultured inside the thermoresponsive polymer solution for periods of up to 21 days. Using this system, cell migration could be assessed by light microscopy at fixed time intervals. At the end of the culture period, the gel could be removed from all grafts and immunohistochemistry analysis showed that endothelial cells were able to maintain confluence, viability, and junctional integrity. Some problems were encountered when using the thermoresponsive cell culture system. These were mostly structural inconsistencies during the sol-to-gel transition phase that resulted in the formation of tiny bubbles in the matrix. Additionally, areas with different viscosity resulted in optical distortions showing up as folds throughout the matrix which can persist even after several cycles of culture medium exchange. These effects had impact on the imaging quality but did not affect the viability of the explant tissue. Conclusion: This study proves that temperature-reversible hydrogel is a very useful matrix for studying in vitro corneal endothelial cell migration from explant grafts and allows for subsequent biological investigation after gel removal. |
| فهرسة مساهمة: | Keywords: Polymer matrix; corneal transplantation; endothelial cell culture; gel matrix |
| تواريخ الأحداث: | Date Created: 20200731 Date Completed: 20211203 Latest Revision: 20211214 |
| رمز التحديث: | 20221213 |
| DOI: | 10.1080/02713683.2020.1798468 |
| PMID: | 32727221 |
| قاعدة البيانات: | MEDLINE |
PDF full text from Publisher | تدمد: | 1460-2202 |
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| DOI: | 10.1080/02713683.2020.1798468 |