To assess tumor-initiating capabilities, NOD.CB17-Prkdcscid mice (Jackson Laboratory; RRID: IMSR_JAX:001303) aged between 8 and 12 weeks old were injected subcutaneously with decreasing numbers of sorted C4-2 cells (100 to 10 cells) under Wake Forest School of Medicine IACUC Approval #A15-221 in accordance with relevant guidelines and regulations. Two tumors were injected per mouse. After 30 days, tumors were removed and weighed. For serial tumor initiation, tumors were dissociated using the Miltenyi tumor dissociation kit run on the Miltenyi gentleMACS Octo Dissociator. Resulting cells were again bead isolated and injected into a second group of mice at 10 cells and allowed to grow 30 days before tumor removal.
Do you have any questions about this protocol?
Post your question to gather feedback from the community. We will also invite the authors of this article to respond.