A method to culture human alveolar rhabdomyosarcoma cell lines as rhabdospheres demonstrates an enrichment in stemness and Notch signaling.

Katherine K Slemmons, Michael D Deel, Yi-Tzu Lin, Kristianne M Oristian, Nina Kuprasertkul, Katia C Genadry, Po-Han Chen, Jen-Tsan Ashley Chi, Corinne M Linardic

Biology open 2021 Feb 09

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The development of three-dimensional cell culture techniques has allowed cancer researchers to study the stemness properties of cancer cells in in vitro culture. However, a method to grow PAX3-FOXO1 fusion-positive rhabdomyosarcoma (FP-RMS), an aggressive soft tissue sarcoma of childhood, has to date not been reported, hampering efforts to identify the dysregulated signaling pathways that underlie FP-RMS stemness. Here, we first examine the expression of canonical stem cell markers in human RMS tumors and cell lines. We then describe a method to grow FP-RMS cell lines as rhabdospheres and demonstrate that these spheres are enriched in expression of canonical stemness factors as well as Notch signaling components. Specifically, FP-RMS rhabdospheres have increased expression of SOX2, POU5F1 (OCT4), and NANOG, and several receptors and transcriptional regulators in the Notch signaling pathway. FP-RMS rhabdospheres also exhibit functional stemness characteristics including multipotency, increased tumorigenicity in vivo, and chemoresistance. This method provides a novel practical tool to support research into FP-RMS stemness and chemoresistance signaling mechanisms. © 2021. Published by The Company of Biologists Ltd.

Citation

Katherine K Slemmons, Michael D Deel, Yi-Tzu Lin, Kristianne M Oristian, Nina Kuprasertkul, Katia C Genadry, Po-Han Chen, Jen-Tsan Ashley Chi, Corinne M Linardic. A method to culture human alveolar rhabdomyosarcoma cell lines as rhabdospheres demonstrates an enrichment in stemness and Notch signaling. Biology open. 2021 Feb 09;10(2)