Abstract
Objective: To establish an immortalized cell line of endometrium through transfection with the plasmid psv3-neo encoding for the sv40 large T antigen and the neomycin gene.
Design: Basic study.
Materials/Methods: Tissue specimens used in this study were obtained from three women who had been given diagnostic curettage before laparascropy. Endometrium tissue was minced into small pieces and treated with collagenase to dissociate epithelial glands from stromal cells, which were further separated by centrifugation. The purity of epithelial cells in primary cultures were propagated and maintained in DMEM. The primary cultures were transfected with psv3 and maintained without FBS for 4–6 hours when it confluenced 80%. After 48 hours cultures were passaged at a split ratio of 1:5. Subcultures were maintained in DMEM containing 10% FBS and the medium was changed at 3-day intervals.
Results: Three discernible clones were observed 5 weeks after transfection. The clones have been maintained in culture for over 12 weeks and was still proliferating without any noticeable change in the biological properties investigated.
Conclusions: These clones may be from immortalized cell lines and the cell lines may be an interesting tool for the study of the pathophysiology of endometriosis. We detected the clones through immunohistochemical, chromosome analysis and molecular biology methods.
Design: Basic study.
Materials/Methods: Tissue specimens used in this study were obtained from three women who had been given diagnostic curettage before laparascropy. Endometrium tissue was minced into small pieces and treated with collagenase to dissociate epithelial glands from stromal cells, which were further separated by centrifugation. The purity of epithelial cells in primary cultures were propagated and maintained in DMEM. The primary cultures were transfected with psv3 and maintained without FBS for 4–6 hours when it confluenced 80%. After 48 hours cultures were passaged at a split ratio of 1:5. Subcultures were maintained in DMEM containing 10% FBS and the medium was changed at 3-day intervals.
Results: Three discernible clones were observed 5 weeks after transfection. The clones have been maintained in culture for over 12 weeks and was still proliferating without any noticeable change in the biological properties investigated.
Conclusions: These clones may be from immortalized cell lines and the cell lines may be an interesting tool for the study of the pathophysiology of endometriosis. We detected the clones through immunohistochemical, chromosome analysis and molecular biology methods.
| Original language | Undefined/Unknown |
|---|---|
| Journal | Fertility and Sterility |
| DOIs | |
| Publication status | Published - 2002 |