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In an original constructed example, seed $300000$ biological cells in $6\mathrm{mL}$ of cell culture medium. Suppose the recoverable count becomes $1200000$ before the next subculture. Transfer a well-mixed quarter of that harvest into a new vessel and bring its final volume to $6\mathrm{mL}$. Assume complete recovery, no loss during transfer and no division during handling. The selected portion containsThe other $900000$ biological cells remain outside the tracked vessel. The lower count after transfer is selection of a portion, not evidence of death. Adding fresh cell culture medium lowers the number density further without changing the selected count.

If the next interval again changes $300000$ to $1200000$, the two growth intervals contribute $2+2=4$ net cell line population doublings. At the second harvest there has been just one intervening passage. Another quarter-transfer would add another passage without adding a growth interval. Thus neither passage number nor the logarithm of the final count divided by the very first seed count records all growth when portions are removed.

For positive recoverable counts $N_0,N_1$ bracketing one growth interval, define its net cell line population doublings byThe formula counts how many factors of two produce the observed net factor. For exactly two divisions per original biological cell, with all daughters recovered, $N_1=2^2{N}_0$, giving $d=2$. Death or loss of biological cells makes this a net estimate of division.^[ref]

Under constant-rate exponential growth, $N(t)=N_0{e}^{rt}$, a growth interval of duration $\Delta t$ givesFor the constructed fourfold increase over $48\mathrm{h}$, $d=2$ and the model doubling time is $24\mathrm{h}$. The proof follows by taking the natural logarithm and solving $e^{r{t}_2}=2$. This is a fitted property of that interval and model: it is not a universal HeLa doubling time, nor the interval between two divisions of each individual biological cell. Across successive growth intervals, add their $d$ values; keep transfer losses separate as in cell culture passage accounting.

An immortalised cell line can apparently sustain unlimited cell line population doublings under suitable conditions. Immortality describes lineage reproduction, not individual biological cell survival.^[ref]