|Year : 1981 | Volume
| Issue : 3 | Page : 151-152
Lens organ culture
AG Chandorkar, MV Albal, PM Bulakh, MP Muley
Dr. V.M. Medical College, Solapur, India
A G Chandorkar
Department of Pharmacology, Dr. V.M Medical College, Solapur-413 003
Source of Support: None, Conflict of Interest: None
|How to cite this article:|
Chandorkar A G, Albal M V, Bulakh P M, Muley M P. Lens organ culture. Indian J Ophthalmol 1981;29:151-2
The incubation of the total lens either in a continuous perfusion fluid or immersed in tissue culture media constitutes a form of organ culture. Lens culture studies, in which the lens was maintained viable for 1 to 3 days, have been reported using either direct lens immersion in tissue culture media,,, or in a perfusion system,,. Almost all reported methods use lenses of rabbits. Hence, the present study was undertaken to evaluate the possibility of using lenses from human, goats, rats and rabbits for organ culture and to study the effect of varying, pH, temperature, media, anatomical positions and submersion or exposure to air.
| Materials and methods|| |
Lenses obtained from the human patients, goats, rabbits and rats were used for the study. Human lenses collected from operation theatre, were kept in saline and transported in ice-box. While fresh goat lens were obtained from the frozen eye balls transported immediately from the slaughter house. Rat and rabbit lenses were dissected out by the method described by Haddad et al for rabbits.
Lenses were carefully placed on a sterile tissue culture dish having a dark blue coloured nylon net, and were maintained either in tissue culture media T.C. 199, with or without either 20% human, rabbit or goat serum respectively;
or in a saline solution isotonic with aqueous. Rat lenses were maintained in isotonic saline and T.C. 199 without any serum. Experiments were carried out at either the pH of 7.2 or 7.8 and at room temperature (27° -29° C) or 37° C in an incubator.
Transparancy was measured by observing number and the characteristics of the squares seen through the lens. Lenses were observed for development of generalised haziness or opacities, intumescence or swelling, disruption and other morphological changes and weight gain. Changes were noted visually and recorded photographically.
Effect of partial and complete submersion of the lens in media fluid and placing on its anterior or posterior surface on the nylon grid was also studied.
| Observations|| |
Lenses from all the four species could be maintained in either T.C. 199 with or without the addition of 20% serum or in isotonic saline for 3-7 days. Qualitatively similar and parellel pattern of change was seen.
The earliest change observed in a normal lens was an opacity starting peripherally and sometimes having a central cataract as well seen at the end of 30-40 hrs. Lens then showed swelling or intumuscence, as confirmed by thickening and distortion of lines of squares seen through the lens, with broadening of the squares and reduction in their number. As the opacity progressed (48-72 hrs) the lens became gradually transluscent and hazy. By the end of 72 hrs the lens developed full intumescence with a gain in weight of 100-500 mg and became completely opaque. It later started to disrupt and disintegrate by 96-120 hrs. The experiments were terminated by removing the lens from media fluid.
Rat lenses were least viable of the three animal species (3-4 days), rabbit lenses were more viable (4-5 days) and goat lenses were the most viable (5-7 days).
Lenses could be maintained at either pH of 7.2 or 7.8 and in both media or temperature without any significant effect on this pattern. However, if kept on anterior surface down or if submerged completely in culture media or saline, it hastened the above process of development of opacities by 2-3 days.
| Discussion|| |
Our results indicate that lenses from other animal species like goats and rats in addition to rabbits can be used for lens culture. They could also be maintained at room temperature unlike most of the previous reports where they were maintained at 37°C in an incubator. It is interesting to note that when the lens was totally submerged it lost its transparancy much more rapidly than when its anterior surface remained facing the atmosphere. Also when the anterior surface of the lens faced the nylon grid there was a rapid development of opacities and disintegration indicating that either the posterior surface has some pump function different from or may be of an opposite nature to that of anterior surface.
Lens culture can hence be used to study various transport and metabolic phenomenon in the lens and also as an experimental model for study of cataracts and for pharmacological evaluation of the drugs acting on lens specially and eye in general.
| Summary|| |
Lenses of various species were maintained in either T.C. 199 media or in a saline solution made isotonic with aqueous humour. Effect of pH, temperature, media, anatomical position, submersion or exposure to air was studied.
It was noted that the lenses can be maintained for 3-7 days in pH ranges of 7.2 to 7.8 in both the media, at 28-29°C (R.T.) or at 37°C. They should be placed on posterior surface and should not be submerged completely. The earliest changes start by 30-40 hrs. as an initial haziness, or peripheral opacities and which are followed by central cataract. Lenses swell by the end of 48-72 hrs. and usually disrupt and disintegrate by 96-120 hrs. These changes can be recorded visually as well as photographically.
| Acknowledgement|| |
The authors thank the Dean, Dr. V.M. Medical College, Solapur for the facilities given to undertake this work.
The work was supported by research grant from Shivaji University, Kolhapur.
| References|| |
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Harding, D.V., Rothstein, H., and Newman, M.B., 1962, Exp. Eye. Res. 1,457.
Schwartz, B., 1973, Culture of the lens, tissue culture Methods and Applications. New York, Academic Press.
Mathur, S.P., 1976, Acta Sixth Afro-Asian Congress of Ophthalmol., 192-194.
Haddad, H.M., Shore, B., and Fuman M., 1967, Amer. J. Ophthalmol:., 63, 1731.