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  • Murine Hair Follicle Derived Stem Cell Transplantation onto the Cornea Using a Fibrin Carrier.

Murine Hair Follicle Derived Stem Cell Transplantation onto the Cornea Using a Fibrin Carrier.

Bio-protocol (2018-05-20)
Mindy Call, Ewa Anna Meyer, Winston W Kao, Friedrich E Kruse, Ursula Schlӧtzer-Schrehardt
ABSTRACT

The goal of this protocol is to establish a procedure for cultivating stem cells on a fibrin carrier to allow for eventual transplantation to the eye. The ability to transfer stem cells to a patient is critical for treatment for a variety of disorders and wound repair. We took hair follicle stem cells from the vibrissae of transgenic mice expressing a dual reporter gene under the control of the Tet-on system and the keratin 12 promoter (Meyer- Blazejewska et al., 2011 ). A clonal growth assay was performed to enrich for stem cells. Once holoclones formed they were transferred onto a fibrin carrier and cultivated to obtain a confluent epithelial cell layer. Limbal stem cell deficient (LSCD) mice were used as the transplant recipient in order to test for successful grafting and eventual differentiation into a corneal epithelial phenotype.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sodium phosphate monobasic, BioXtra, ≥99.0%
Sigma-Aldrich
Sodium phosphate dibasic, BioXtra, ≥99.0%
Sigma-Aldrich
Glycerol, BioXtra, ≥99% (GC)