Primary explant culture and collagen I substrate enhances corneal endothelial cell morphology

Autor: Carlos-Alberto Rodriguez-Barrientos, María-Dolores Montalvo-Parra, Guillermo-Isaac Guerrero-Ramírez, Judith Zavala, Jorge E. Valdez-García, Victor Trevino
Jazyk: angličtina
Rok vydání: 2018
Předmět:
0301 basic medicine
Corneal endothelium
Morphology (linguistics)
Immunocytochemistry
Cell Culture Techniques
Gene Expression
lcsh:Medicine
engineering.material
Data Note
General Biochemistry
Genetics and Molecular Biology
Collagen Type I
03 medical and health sciences
0302 clinical medicine
Coating
Glypicans
Antigens
CD
Humans
Primary cell culture
lcsh:Science (General)
lcsh:QH301-705.5
Cells
Cultured
Cell Proliferation
Matrigel
Chemistry
Reverse Transcriptase Polymerase Chain Reaction
Endothelium
Corneal
lcsh:R
Substrate (chemistry)
Endothelial Cells
General Medicine
Collagen I
Molecular biology
Immunohistochemistry
Drug Combinations
030104 developmental biology
lcsh:Biology (General)
Cell culture
030221 ophthalmology & optometry
engineering
Zonula Occludens-1 Protein
cardiovascular system
Proteoglycans
Collagen
Laminin
Sodium-Potassium-Exchanging ATPase
Explant culture
lcsh:Q1-390
Zdroj: BMC Research Notes, Vol 11, Iss 1, Pp 1-3 (2018)
BMC Research Notes
ISSN: 1756-0500
DOI: 10.1186/s13104-018-3174-3
Popis: Objectives Corneal endothelial cell (CEC) isolation and harvest aim to produce engineered grafts to solve donor corneal tissue shortage. To yield high amounts of CEC maintaining morphological and molecular characteristics, several isolation and culture conditions are reported. Here, we combined direct explant culture, with three different coating conditions and a two-step media approach to compare confluence efficiency, morphology, and specific molecular markers expression. Data description Confluence was reached after 2 weeks in the three coating conditions (Matrigel, collagen I, and in uncoated plates) using a two-step approach (proliferative medium without pituitary extract, followed by stabilizer basal medium). Na/K-ATPase and GPC4 markers were detected by immunocytochemistry while GPC4, CD200, and TJP1 by RT-PCR in the three CEC coating culture conditions. CEC in proliferative medium showed spindle morphology in the three conditions. Polygonal morphology was seen in CEC cultures using basal medium under uncoated and collagen I coated plates. CEC cultured in Matrigel-coated plates remained with spindle morphology in basal medium.
Databáze: OpenAIRE
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