PA5 hOMCs growth and metabolic profile in cell cultures using
Plastic and Plastic Plus microcarriers
A method previously developed by Santos et al 2011 was adopted to
performed PA5 hOMCs microcarrier cell culture in spinner flasks. A total
working volume of 80 mL in 100 mL Belco spinner flasks was used with
50% of media change every two days, for a total of 7 days. An initial
seeding density of 6000 cells/cm2 was used, which is
equivalent to 3.4x104 cells/mL. Figure 3 A shows that
both Plastic and Plastic Plus microcarrier cultures experienced a
decrease in cell concentration after day 1 from
3.4±0.03x104 cells/mL to 1.4±0.3x104cells/mL and 9.4±1.3x103 cells/mL, respectively. Cell
number started increasing after day 3. On day 7, the concentration of
cells in cultures employing Plastic microcarriers was
1.4±1.1x105 cells/mL and for Plastic Plus
microcarriers was 4.9±1.8x104 cells/mL (statistically
significant, p<0.05). Cell viability remained above 90%
(Figure 3 A) throughout the culture period. Metabolic analysis of
glucose, lactate and ammonia revealed similar concentration of these
nutrient and metabolites for both conditions (Figure 3 B-D). On day 7,
glucose concentration was of 10.5±1.7 mM and 11.5±1.6 mM, while lactate
concentration was of 7.3±2.3 mM to 5.2±2.5 mM, for PA5 hOMCs grown on
Plastic and Plastic Plus, respectively. The concentration of ammonia was
of 0.7 mM for both conditions. Considering an exponential phase starting
on day 3, specific consumption of glucose, specific production of
lactate and ammonia, as well as yield of lactate per glucose were
calculated. PA5 hOMCs grown of Plastic Plus led to a specific glucose
consumption rate of 24.4±2.3
pmol.day-1cell-1, similar to the one
measure for PA5 hOMCs grown on Plastic which was 20.4±4.7
pmol.day-1cell-1. Specific
production rates for lactate and ammonia was 47.9±6.3
pmol.day-1cell-1 and 0.6±0.1
pmol.day-1cell-1, respectively for
PA5 hOMCs grown on Plastic Plus. These results were on average 2 times
higher than the specific production rates observed for PA5 hOMCs grown
on Plastic microcarriers (24.6±7.2
pmol.day-1cell-1 and 0.3±0.05
pmol.day-1cell-1 for lactate and
ammonia, respectively). The yield of lactate per glucose was 1.4±0.4 and
2.0±0.3 for PA5 hOMCs grown on Plastic and Plastic Plus microcarriers,
respectively.