Table 5 : T cell behaviour in MO, MH and SH bottom sediments and their capacity for induced terminal differentiation.


Sample no. OCB dose (mg) Counting time (h) T cells (D1+D2) Cell divisions (A/S) Slow-fast cycling MAT cells (D2) ITD cysts ITD cysts % Refractory to ITD

5a 5 12 5.2 1 (A) -- D21 4.37 84 --
5b(1) -- 18 7.6 2 (A) fast D21-2 4.71 62 D13
5c(1) -- 36 15.2 5 (A) fast D21-5 15.21 100 --
5d -- 48 18.8 6 (A) -- D21-6 17.86 95 --
5e(2) 10 48 13.2 4 (A) intermediate D21-4 10.56 80 D14
5f(2) -- 60 15.6 5 (A) -- D21-5 n.d. -- --
5g(3) 15 48 6.0 1 (S) slow ISH 12.08 200 --
5h -- 72 17.2 1(S)+2(A) -- D22-3 10.32 80 --

Cell fractions in 105 units; S, symmetric division; A, asymmetric division Parallel subcultures were started with a low amoebic inoculum of about ¼ of the standard amoebic inoculum (2.60x105 T cells).
OCB used in these experiments are metabolically different from OCB in the table 3 and 4. Samples 5a-5d receive the standard OCB dose of 5 mg, samples 5e-5f the doubled and samples 5g-5h the
tripled bacterial dose. All cultures were evaluated only once.
In cultures with 5mg OCB, T cells end current cell cycle and divide at the end of the initiation phase INI at about t4. Thereafter, the t-SRL lines of the sample 5b/5c proliferate by fast cycling (AGT:6.2-7.2 h).
In cultures with the doubled bacterial dose of 10 mg (sample 5e/5f) the t-SRL line proliferates with moderate AGT values (AGT11-12). The strong hypoxia which developed in culture sediments with 15mg
OCB (samples 5g-5h) converted T cell progeny into symmetric cell fate. T cells converted to ISH cells. ISH cells progress cell cycle slowly and remain arrested at checkpoint for T cell re-conversion. Between
t48 and t72 ISH cells return to asymmetric cell fate and AGT11.
ISH and MAT (D2) cells are capable for induced strong hypoxic terminal differentiation. Both cell types give rise in strong hypoxic OCB sediments and AaEM encystment medium to ITD cysts. Self-renewing
T cells (D1) cells may be induced only in the early G1 phase (samples 5c, 5d) so long as they are in a state of double potency and not mitoticly determined. When mitoticly determined they become
refractory to encystment stimuli (samples 5b, 5e).

Vladimir F Niculescu Stem Cell Biology and Research  2015 2:2DOI : 10.7243/2054-717X-2-2