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Fig. 2. AHNPs avoid immortalizing mutations, and exhibit mitogen- and
telomerase-dependent growth. (A) Cultured AHNPs express major
growth regulatory proteins longitudinally throughout culture. (B)
Karyotyped AHNPs display normal ploidy and have no gross cytogenetic
malformations. (C) Following growth arrest by an exogenous TERT
inhibitor (EGCG) or growth factor withdrawal cultured cells express
SA-ß-Gal. However, only mitogen-withdrawn (-bFGF) cells lose TERT
expression when evaluated 7 days later. (D) Physiological
(x-irradiation) or chemical inhibitors (apidicolin, EGCG) consistently
increase the fraction of cells expressing SA-ß-Gal. (E)
Application of reversible growth inhibitors yields a significant reduction in
growth rate. AHNPs revert to previous proliferative levels following arrestor
washout. (F) Age-matched AHNPs placed in either basic media (N2) or
media containing EGF or bFGF only (N2E, N2F) enter irreversible growth arrest
compared to defined proliferative conditions (N2EF) and subsequently become
unviable. Data shown for temporal cortex derived cells.
*P<0.05, Student's t-test. Scale bar: 75 µm
in C.
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