Purpose: To optimize the medium and factors required for growth and prevention of cell death in transfected human lens epithelial cells (HLECs) (Andley et al., IOVS 35:3094). To identify factors inhibiting growth. Methods: 100,000 HLECs were grown in 7 different media containing 20%FBS or in MEM containing 0.1-20%FBS. Growth factors were added to MEM containing 0.1%FBS. Factors evaluated included, insulin, IGF-1, IGF-2, ECGS, EGF, FGFa, FGFb, HGF, PDGF-BB, KGF, IL6, SCF, TGFα and TGFβ1, TGFβ2 or TGFβ3. Cells were fed every other day and counted on day 7. Results: HLECs exhibited maximum growth in MEM with 20%FBS, died in MEM alone and exhibited limited growth in MEM supplemented with 0.1%FBS. Addition of any TGFβ at 50pg/ml to MEM+0.1%FBS inhibited growth. Cells cultured in ECGS (pituitary or neural) plus insulin, IGF-1 or IGF-2 exhibited a 3 fold increase over controls that exceeded growth obtained with other growth factor combinations. Addition of TGFβ1 to ECGS+IGF-1 inhibited proliferation. Individual factors that enhanced growth were insulin, IGF-1, IGF-2, ECGS, EGF, PDGF-BB, IL-6, SCF and TGFα. Conclusions: Optimization of medium and serum which stimulate growth and identification of growth factors that prevent death should help define the factors required for the long-term growth and survival of HLECs.
|Journal||Investigative Ophthalmology and Visual Science|
|State||Published - Feb 15 1996|