engleski

Attempts at improving thymic epithelial stem cell isolation protocols

Studies have shown that transplanting thymus cells into patients can help restore the function of the immune system, but the persistent problem is finding the source of specialized stem cells. Thymic epithelial progenitor cells (TEPC) present an alternative source of transplatable cells because they can generate the epithelial subpopulations of the thymus necessary for the maturation of T-lymphocytes. The aim of our study was to isolate thymic epithelial cell precursors from the human thymus and culture them in vitro as a continuous source of cells, as well as developing new methods for their cryostorage. There are several protocols for isolating thymic epithelial cells and a combination of enzymatic digestion and cell enrichment was chosen as a starting point in the attempt at isolation of human TEPC. The efficiency in isolating our desired population of cells was determined using flow cytometry shortly after digestion, as well as 10 days after plating the obtained cell fractions. The major problem in the attempted digestions was the high percentage of thymocytes which obstruct the flow cytometry analysis of much smaller stem cell populations. In addition, the high sensitivity of obtained cells to freezing makes it difficult to study cell samples after a prolonged period of time. Here we present our flow cytometry results before and after cell plating.

TEPC ; isolation protocols ; flow cytometry

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