Table 2 : Growth of the primary culture in sediments with oxygen consuming bacteria.


Time in h (event) ITD cysts Metacystic
amebulae (A)
Vegetative
amoebae (V)
SRP cells
(p-SRL)
MAP cells SRS cells (s-SRL) ATD cysts Cell divisions Ratio A:V

t0 (start) ≥0.1 -- -- -- -- -- -- -- --
Ex-cysment -- ≥0.8 -- -- -- -- -- -- --
t96 (1st counting) -- -- 5.68 ≥0.8 ≥4.8 -- -- 6 1:7
t120 (2nd counting) -- -- 5.76 -- ≥4.8 ≥0.8 1.7 6+2 1:7

Cell fractions in 106 units. Approximately 0.1x106 ITD cysts induced by AaEM encystment medium were added to an Aa(Sm) culture of 5 ml medium and 5 mg metabolically
repressed OCB. To minimize oxidative stress by stirring, counting and re-sedimentation (SARC), samples were counted only toward the end of the growth phase (t96). After
ex-cystment ≥0.8x106 metacystic amebulae (A cells) started the primary p-SRL by A/P conversion. Amoebic population increased by arithmetic progression and asymmetric
cell division producing non-identical D1 and D2 daughter cells. At t96 1/7 of the population (≥0.8x106) belongs to the cycling self-renewing p-SRL line in the sixth generation
(D16 cells). The other six parts (≥4.8x106 cells) are MAT cells (D21-6 cells) accumulated in the quiescent stem cell reservoir. They remain mitoticly repressed until the end of
the culture, yet retain phagocytic activity.
SARC/t96 leads p-SRL line to P/S-conversion, starting the secondary s-SRL stem cell line. The s-SRL line remains numerically unchanged (≥0.8x106 cells). During the time
t=96 until t=120 it goes through two cell cycles. Two additional generations of MAS cells (D27-8) give rise to ATD cysts by autonomous terminal differentiation. The primary
culture becomes heterogeneous. At t=120 it contains ≥4.8x106 primary MAP cells (D21-6 cells), ≥0.8x106 self renewing secondary SRS cells (D18 cells) and 1.7x106 ATD cysts
(encysted MAS, D27-8 cells). The primary p-SRL line proliferates by AGT≤15 while the secondary s-SRL line by AGT6.

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