Development and Survival of Human Ovarian Cells in Chitosan Hydrogel Micro-Bioreactor.

Background and Objectives: To test the long-term ability of human ovarian cortex cells to develop in unconventional culture conditions. Materials and Methods. Ovarian cortex cells from fetuses aged 23 to 39 weeks gestation were cultured for 90 days in hollow chitosan hydrogel micro-bioreactors and concurrently in traditional wells. Various cell-type counts were considered. Results: With intact follicles as a denominator, the percentage of growing intact follicles at Day 0 varied widely between ovaries (0 to 31.7%). This percentage tended to increase or stay relatively constant in bioreactor as in control cultures; it tended more toward an increase over time in bioreactor vs. control cultures. Modeled percentages showed differences (though not significant) in favor of bioreactor cultures (16.12% difference at D50 but only 0.12% difference at D90). With all follicles present as a denominator, the percentage of growing primary and secondary follicles at D0 varied widely between ovaries (0 to 29.3%). This percentage tended to increase over time in bioreactor cultures but to decrease in control cultures. Modeled percentages showed significant differences in favor of bioreactor cultures (8.9% difference at D50 and 11.1% difference at D90). At D50 and D90, there were only few and sparse apoptotic cells in bioreactor cultures vs. no apoptotic cells in control cultures. Conclusions: Over three months, bioreactor folliculogenesis outperformed slightly traditional culture. This is an interesting perspective for follicle preservation and long-term toxicological studies.

Preliminary study of the cultivation and growth characteristics of human salivary gland epithelial cells / 实用口腔医学杂志

Objective:To establish a culture method of human salivary gland epithelial cells and to study their growth characteristics in vitro.Methods:Tissue explant technic was employed to culture human salivary gland epithelial cells in serum free medium (SFM),1∶1 DMEM/F12 and 1∶1 DMEM/F12 containing 25 ml/L fetal boven serum (FBS) respectively.The morphology of the cultured cells was observed by phase contract microscope.The cell growth was studied by cell counting.The cells were identified by HE staining,PAS staining and SP staining.Results:Growth of human salivary gland epithelial cells was observed in primary culture in the three kinds of medium in all 10 cases. The cultured cells were epidermoid, positive for cytokeratin, negative for Vimentin and positive for PAS staining. The cells in SFM could be subcultured for five passages,while only for one passage in the other two kinds of medium.Conclusion:SFM is superior to serum free 1∶1 DMEM/F12 or 1∶1 DMEM/F12 containing 25 ml/L FBS for the culture of human salivary gland epithelial cells.