engleski

Quality Control of Cultivated Limbal Epithelial Stem Cell For Transplantation

Cultivation of limbal stem cells (LSC) is a well-established method of cellular therapy for patients with limbal epithelial stem cell deficiency caused by chemical burns, thermal injuries or chronic immune inflammatory diseases. Limbal cells were obtained from ocular biopsies cultivated on a feeder-layer made from 3T3 cells. This cell therapy was classified as advanced therapy medicinal products (ATMPs).In the Laboratory of Tissue Engineering in microbiological cabinet (which is clean room class A) were we prepared limbal stem cell grafts. With methods of flow cytometry it is possible to detect anti-feeder layer positive cells, stem cells and differentiation cells.Haematoxylin and eosin stained histological images showing firstly cell morphology and secondly stratification of limbal grafts. Immunohistochemistry analysis detected positive staining of stem cells (p63α), Ki67 and CK3. Live-cell imaging of human LSC were obtained in collaboration with Ruđer Bošković Institute, Zagreb. The cell forms a spindle, a molecular machine responsible for equal distribution of chromosomes between the daughter cells. It is recommended to use reagents, growth medium and water certified to contain endotoxin in minimal concentration. Cultivated limbal epithelial transplantation (CLET) has shown to be an effective therapy for LSCD, with clinical trials reporting an average success rate of 70 to 80%

limbal stem cells

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