METHOD FOR ACCELERATING THE GROWTH OF ANCHORAGE-DEPENDANT CELLS IN NON-STATIONARY CULTURE CONDITIONS.

摘要

Human haematopoietic stem cells extracted from the umbilical cord were grown under non-stationary culture conditions. The culture environment consisted on subjecting the cells to different rotational fields of laminar speed by means of a novel rotating equipment. The growth kinetics of each non-stationary environment was compared against the common kinetics under stationary environments for assessing the cell differentiation due to the culture conditions. The expression levels of the marker CD34 were compared by the polymerase chain reaction with reverse transcription (RT-PCR) and agarose gel electrophoresis. The results suggest that the cells cultured at 3 rpm and at radial distances of from about 6 cm to about 9.5 cm have a shorter latency phase and produce cell concentrations 46% an 66% higher, respectively, than those of the stationary culture during the 10th day of the culture period. The rotation at 3 rpm and at the radial distances of 6.25 cm and 9.5 cm has been reduced, in the duplication time during the exponential phase and regarding the stationary culture, nearly 19% and 18%, respectively. At 5 rpm of agitation, the cells had a latency period 100% shorter and produced 57% and 60% more cells with regard to the control at 6.5 cm and 9.5 cm, respectively.