Cellular Medicine Research Breakthroughs in the Control of Diseases:
Report on Scientific Research
S. Netke, Ph.D., V Ivanov , MD. Ph.D., W. Roomi, Ph. D., A Niedzwiecki,
Ph. D., Matthias Rath, M.D.
Presented at: 21st Century Medical Advances in the Treatment of Cancer
and Aging, Miami , Fl, March 22-24, 2003; Published in: Conference Proceedings
Method: Normal human dermal fibroblasts (NHDF ) were grown to confluence
on insert of “Improved Matrigel Invasion Chamber” using DMEM
media. The media was withdrawn and the insert and the well were washed
with sterile PBS. Human breast cancer cells (5 X 104) suspended in Liebovitz’s
media were seeded on the insert. Various supplements were added to the
Liebovitz’s media and 750ul and 200ul media were added to the well
and on the insert respectively. The chambers were incubated in free gas
exchange incubator for 22 hours. After incubation the media was withdrawn
and the cells on the upper surface of insert were wiped off with cotton
swab. The cells on the lower surface were stained.
Results: Supplementation of media with 100 uM ascorbic acid (AA) reduced
the invasion by 36%. Supplementation with AA+ with 140uM proline (P) further
reduced the invasion by 47%. Using 400 uM lysine on the top of AA and
P in the supplement caused the invasion to be reduced to 67%. Further
studies with graded levels of Lysine with AA and P showed that there was
linear response to lysine up to 800uM level.