Figure 1.

Measurement of OCM proliferation rates. (A) OCMs were transduced with a lentivirus expressing a GFP-tagged histone 2B (GFP-H2B) to generate cells with fluorescent nuclei. Time-lapse microscopy was used to measure fluorescent object count to infer nuclear count, as a proxy for cell number, over 120 h. The inverse gradient of the linear portion of a log₂ transformation of the fluorescent object count, normalised to t = 0, was used to determine culture doubling time. (B) Doubling times of the present cohort of 16 OCMs and the non-transformed FNE1 cell line ranked from shortest to longest duration. Mean and SD determined from a minimum of two independent experiments. (C) Comparison of mean doubling time from (B) with Nelson et al., 2020 for the 11 OCMs included in both analyses (note, for drug screening purposes the present cohort excludes OCMs with relatively slow proliferation rates and includes novel OCMs not available at the time of the prior analysis). A y = x line is shown for illustrative purposes. (D, E) Comparison of doubling time with aneuploidy score (D) and structural score (E) from shallow, single-cell, whole-genome sequencing (both datasets from Nelson et al., 2020). Linear regression is shown and Pearson's r is used to measure correlation. Two-tailed P value, ** P ≤ 0.01. Note that the culture established from the third biopsy from patient 64 was further separated into EpCAM-negative (64-3-) and EpCAM-positive (64-3+) cells, with only OCM.64-3- included in the current analysis (15). See also Supplementary Figure S1.