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Table 1 Characteristics of primary, immortalized and tumorigenic cells in the panel

From: Increased OXPHOS activity precedes rise in glycolytic rate in H-RasV12/E1A transformed fibroblasts that develop a Warburg phenotype

cell population

abbreviation

description

cell diameter (μm)

cell volume (pL)

number of colonies in soft agar

tumor size (mm2) in BALB/c nu/nu

cell doubling time (h)

Primary MEF

Prim-MEF

Isolated from 12–14.5 day-old embyos

16.3 ± 1.0 (3)

2.6 ± 0.5 (3)

none

n.a.

20.5 ± 0.7 (6)

Immortalized MEF (spontaneous)

Imm-MEF

Senescence bypass (3T3 passaging)

16.5 ± 0.7 (3)

2.7 ± 0.3 (3)

none

n.a.

24.1 ± 1.1 (6)

Immortalized MEF (TBX2)

TBX2-MEF

Retroviral transducton TBX2 gene

15.5 ± 0.3 (4)

2.1 ± 0.1 (4)

none

n.a.

19.9 ± 0.9 (6)

RasV12/E1A

MEF (low passage)

RAS-LP

Retroviral transducton E1A/RasV12

11.0 ± 1.0 ** (4)

0.82 ± 0.2 ** (4)

131 ± 68 (5)

131 ± 62 (9)

16.8 ± 1.6 # (6)

RasV12/E1A MEF (high passage)

RAS-HP

Retroviral transducton E1A/RasV12

10.1 ± 0.4 *** (4)

0.58 ± 0.1 *** (4)

880 ± 164 †† (5)

497 ± 182 † (9)

12.2 ± 0.7*** ‡ (8)

RasV12/E1A MEF (tumor)

Ras-TUM

RAS-HP passage in BALB/c nu/nu mice

10.5 ± 0.3 *** (3)

0.65 ± 0.1 *** (3)

705 ± 146 †† (7)

n.a.

15.0 ± 1.0 ** (7)

  1. Cell diameter, doubling time, and in vitro and in vivo tumorigenic characteristics were analyzed. Values represent average ± SEM (n). Cell diameter and cell doubling time decrease significantly in H-RasV12/E1A-transformed cells. Primary and immortalized cells do not form tumors in vitro or in vivo, whereas tumorigenic capacity of H-RasV12/E1A-transformed cells increases with passage number.
  2. 1For cell diameter analysis, approximately 120 cells per population were analyzed in each of the independent experiments.
  3. 2Soft agar assays were carried out in duplicate in each of the independent experiments.
  4. 3In vivo tumor assays were conducted on a total of nine animals as described in Material and Methods.
  5. 4For cell growth analysis and cell doubling time calculation, at least six independent experiments, each in duplicate, were conducted. Cells were seeded at 128,000 (Prim-MEF, Imm-MEF, TBX2-MEF) or 200,000 (Ras-LP, -HP, -TUM) cells per well of a 6-well plate.
  6. Statistics:
  7. **: p < 0.01; ***: p < 0.001 compared to Prim-MEF (one-way ANOVA/Bonferroni)
  8. ‡: p < 0.05 compared to Ras-LP (one-way ANOVA/Bonferroni)
  9. †: p < 0.05; ††: p < 0.01 compared to Ras-LP (Student's t-test)
  10. #: p < 0.10 for comparison Prim-MEF – Ras-LP (Student's t-test)
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Molecular Cancer

ISSN: 1476-4598