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 The media was withdrawn and the insert and the well were washed with sterile PBS.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 After incubation the media was withdrawn and the cells on the upper surface of insert were wiped off with cotton swab.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.