Related Experiment Videos
Mitogen(s) for endometrial-like cells can be detected in human peritoneal fluid
M Koutsilieris1, L Allaire-Michaud, M Fortier
1Hôpital St-François d'Assise, Département d'Obstétrique-Gynécologie, Université Laval, Québec, Canada.
Objective:
To characterize the presence of mitogen(s) in the peritoneal fluid (PF).
Design:
Aliquots of PF aspirated at laparoscopy were assessed for mitogenic activity by the rate of [3H]-thymidine incorporation into the deoxyribonucleic acid of various cell lines.
Setting:
Peritoneal fluids were obtained from patients having a laparoscopy for the investigation of infertility or pelvic pain or for tubal ligation.
Patients:
Seven women with laparoscopic evidence of endometriosis (stage I and II) and six without evidence of endometriosis.
Intervention:
None.
Main Outcome Measure:
After 24 hours of culture in absence of serum, NIH/3T3 mouse embryo fibroblasts, (KLE) human endometrial adenocarcinoma cells, and primary cultures of rabbit endometrial cells were incubated in the presence of appropriate amounts of PF or bovine serum albumin (control) for 22 hours before adding the [3H]-thymidine for 2 hours.
Results:
Aliquots of PF containing greater than 100 micrograms/mL total protein concentration stimulated the proliferation of all cell types. This mitogenic effect was dose-dependent and was greatest on endometrial-like cells. The mitogenic activity was fully recovered after adsorption on dextran-coated charcoal and was sensitive to tryptic digestion and to heat. It could not be retained on cartridge of C18 silica and was calculated to have a molecular weight greater than 30,000 by gel permeation chromatography.
Conclusions:
Peritoneal fluid from women with or without endometriosis contain growth factor(s) that are proteinaceous in nature and capable of stimulating the proliferation of endometrial-like cells and fibroblasts. These mitogens could play a role in the pathogenesis of endometriosis.
Insights
Peritoneal fluid contains protein-based growth factors that stimulate cell proliferation. These mitogens, found in women with and without endometriosis, may contribute to the development of endometriosis.
Area of Science:
- Gynecology
- Cell Biology
- Biochemistry
Background:
- Endometriosis is a condition where uterine lining tissue grows outside the uterus.
- The peritoneal fluid (PF) environment may influence the development and progression of endometriosis.
Purpose of the Study:
- To identify and characterize mitogens present in human peritoneal fluid.
- To investigate the potential role of these mitogens in the pathogenesis of endometriosis.
Main Methods:
- Peritoneal fluid samples were collected from women undergoing laparoscopy.
- Mitogenic activity was assessed using [3H]-thymidine incorporation assays with various cell lines, including mouse fibroblasts and human endometrial cells.
- Physicochemical properties of the mitogens were determined through adsorption, enzymatic digestion, heat treatment, and chromatography.
Main Results:
- Peritoneal fluid with protein concentration >100 µg/mL demonstrated dose-dependent mitogenic activity.
- Endometrial-like cells exhibited the greatest proliferation in response to PF.
- The mitogenic factors were identified as heat-stable, proteinaceous substances with a molecular weight >30,000 Da, sensitive to tryptic digestion.
Conclusions:
- Human peritoneal fluid contains proteinaceous mitogens capable of stimulating fibroblast and endometrial cell proliferation.
- These mitogens may play a significant role in the pathogenesis of endometriosis.
- Further research into these growth factors could offer new therapeutic targets for endometriosis.