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. 2021 Jun 6;10(6):1413. doi: 10.3390/cells10061413

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Procedure of isolation, immortalization and subcloning of urine-derived kidney epithelial cells. Fresh urine is cultured in a specific medium that selects for specific types of kidney epithelial cells at 37 °C. Immortalization is performed by use of SV40 and the hTERT gene, upon which antibiotic selection is used to isolate the successfully immortalized cells. Clonal colonies are selected, in order to isolate and expand specific cell clones that will be characterized by biomarker expression and stored for cryopreservation.