| Publication number |
CLPUB00039 |
|---|
| Authors |
Reddan J.R., Lindemann C.B., Hitt A.L., Bagchi M., Raphtis E.M., Pena J.T., Dziedzic D.C. |
|---|
| Title |
Generation of two non-transfected human lens cell lines. |
|---|
| Citation |
Invest. Ophthalmol. Vis. Sci. 40:S970-S970(1999) |
|---|
| Web pages |
https://eurekamag.com/research/034/979/034979197.php |
|---|
| Abstract |
Purpose: To establish long-term lines of non-transfected human lens
epithelial cells (HLECs). In all previous studies from numerous
laboratories, non-tnasfected HLECs complete at most 3.4 passages and
undergo senescence. The valuable information that has been discovered
using animal models needs to be validated on a human model system.
Methods: A novel method was developed wherein capsule-epithelial explants
were placed on microporous membranes in a 1:4 mixture of keratinocyte
growth medium and M199 over a 1:1:1 mixture of fetal bovine, rabbit and
horse sera. It was our rationale that the basal diffusion of 100% serum
through the membrane onto the explant would mimic a wound-healing
environment and possibly stimulate stem cell division.
Results: Cells were cultured for 4 passages (~5 months) over the 100%
serum mixture before growth became vigorous. After this time, the cells
showed continued growth even when transferred to conventional culture
dishes. The cells exhibit an epithelial morphology and are currently at
passage 20, population doubling level 50. One of the lines is primarily
diploid. The cells grow in response to insulin, IGF1 and ECGS. Cells form
colonies when plated at low density and can he cryopreserved and recovered.
Conclusions: Using a new and unique method to stimulate growth, we have
established two stable lines of HLECs. This technique may permit
establishment of cell lines from cataractous lenses.
|
|---|
| Cell lines |
CVCL_S908; FHL124 |
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