MAINTENANCE OF HUMAN LENS EPITHELIAL CELLS IN TISSUE CULTURE

Human lens epithelial (HLE) cell cultures arewere distributed into 3 groups of normal, control
advantageous forstudying the etiology of cataract andandtreated. Effect of Ocimum sanctum (70-280 mg
may serve as a model forscreening anti-cataractml) was studiedon the development of the opacity and
agents. In the present study, anteriorcapsule explantson related biochemicalparameters. Oxidative stress
of epithelial cells from a 25 year old woman,obtainedwas induced by incorporatingsodium selenite (100 mM)
from the National Eye Bank, Dr. R.P. Centre,in TC-199 media. Lenses were incubatedin a 24 well
AIIMS,seven hours after the death, were cultured inFalcon plate at 37o C with 5% CO2 for 24
Dulbecco's ModifiedEagle's Medium supplemented withh.Thereafter, lenses were washed, weighed and
20% fetal calf serum. Lenscapsule in four pieces wasprocessed forestimation of glutathione,
stuck to the surface of a petridishwith epithelium facingmalondialdehyde, soluble and insolubleprotein. Enzyme
upwards. It was first incubated for 30minutes in a CO2activities viz. catalase, glutathione peroxidaseand
incubator with only few drops of medium on aside forglutathione-S-transferase were measured in the lenses
the proper adherence and subsequently 4 ml ofin aseparate set of similar experiment. For in-vivo
mediumwas poured and incubated further. The primarystudies Ocimumsanctum leaf extract (10 mg/kg) was
cultures wereregularly observed under phase contrastinjected intraperitoneally6h prior to administration of
microscope and themedium was changed twice a0.25 mmole of sodium selenite perpup subcutaneously
week. Cells were trypsinized andsubcultured on theon the 10th postnatal day. Pups of bothgroups were
5th day. Cell outgrowth was observed fromeverysacrificed on day 1, 2, 3 and 4 post injection,
piece of capsule explant within 72 hours. The primarylenseswere dissected out and processed for protein
cellgrowth continued to spread out during subsequentestimation andSDS PAGE. The opacity was monitored
days ofcultures. The cultures formed monolayer offrom 14th postnatal dayonwards on a slit lamp. Faint
hexagonal cells and became confluent in 7-8 days. Ifopacity was observed at theconcentration of 140 and
not subcultured HLE cellsshowed signs of280 mg/ml in the treated group incomparison to the
degeneration on the 10th day. We couldsuccessfullyopacity in the control group. Incidence ofcataract was
maintain HLE cells upto two passage. Further workis infound to be 100 and 60% in the control and
progress.treatedgroup respectively. Marked difference in SDS
OCIMUM SANCTUM : AN INDIGENOUS PLANTPAGE profile wasnoticed in control and treated groups.
WITHANTICATARACT POTENTIALOcimum sanctum alsopositively modulated antioxidant
The study was carried out to evaluate theparameters. It is concluded thatOcimum sanctum
anti-cataract potentialof aqueous extract of Ocimumshowed anti-cataract potential both in in vivoand in vitro
sanctum (Tulsi) leaves againstselenite inducedexperimental models of cata-ractogenesis.
oxidative stress in vitro and in vivo. Young ratlenses