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L-cysteine improves growth of human peritoneal mesothelial cells in vitro
S D Bird1, M Legge, R J Walker
1Department of Medicine, Faculty of Medicine, University of Otago, Dunedin, New Zealand.
Objective:
To improve the growth characteristics of human peritoneal mesothelial cells (HPMC).
Design:
The effect of commonly used agents, L-cysteine and epidermal growth factor (EGF), added individually ("single") or mixed with hydrocortisone and apo-transferrin ("admixture") in the culture medium (M199) on cultured HPMC, was investigated.
Methods:
Growth agents were added to M199 medium along with 2% fetal bovine serum and L-glutamine. Growth was determined by the analysis of thymidine ([methyl-3H] thymidine) incorporation into deoxyribonucleic acid, total cell protein, and by cell counts. Morphology was assessed by phase contrast light microscopy and scanning electron microscopy.
Results:
HPMC exposed to L-cysteine 0.25 x 10(-3) mol/L (30 micrograms/mL) exhibited significantly improved attachment and growth. Attached cells appeared flat and well spread out shortly after seeding, and produced a tight polygonal monolayer in 14 days, in contrast to the growth of HPMC in control M199 medium, which failed to reach confluence. After an initial lag period in cell growth, EGF (0.01 microgram/mL) produced a greater increase in cell growth than L-cysteine did; however, this was associated with changes in HPMC morphology. During the growth period (14 days), EGF-stimulated HPMC appeared distorted and irregular compared to L-cysteine-treated cells, which had the characteristic tight "cobblestone" appearance.
Conclusion:
L-cysteine improved cell attachment with preservation of the characteristic morphology of HPMC. Epidermal growth factor improved cell growth but produced changes in morphology. The addition of L-cysteine to the culture medium has an important cell growth enhancement role due to the improved cell attachment and cell viability.