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Isolation and Culture of Primary Human Gingival Epithelial Cells using Y-27632
Chapters
Summary November 6th, 2021
Here we present a modified method for the isolation and culture of human gingival epithelial cells by adding the Rock inhibitor, Y-27632, to the traditional method. This method is easier, less time-consuming, enhances stem cell properties, and produces larger numbers of high-potential epithelial cells both for the laboratory and for clinical applications.
Transcript
So general goal of this cell-based protocol is to isolate and encounter Human Gingival Epithelial Cells in order to improve the shortcomings of direct eggplant method.From the new modified endothemic method, we can get larger numbers of primary gingival epithelial cells in a relatively short time, based on our materials and equipment.So human tissue used in this protocol were fresh adult gingival tissues discarded from extractions of impacted teeth in the Maxillofacial Surgery Department according to the guidelines of the Institution's Human Research Ethics Committee.The New Modified Enzymatic Method.Gingival Tissue Pretreatment.Wash the tissues with 75%ethanol for 30 seconds.And then, with washing solution twice for five minutes each.Digestion of Gingival Tissue.Cut the tissues into tiny pieces with two surgical blades.Add 2.5mg/mililitre dispase and collagenase-1 solution into a 1.5 mililitre centrifuge tube.Put the tissue pieces into the 1.5 mililitre centrifuge tube.Incubate for 15-20 minutes in a 37
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