Optimization of The Cell Aggregates Method for Isolation and Purification of Human Granulosa Cells from Follicular Fluid
Journal Title: International Journal of Fertility & Sterility - Year 2020, Vol 13, Issue 4
Abstract
Background: Aspirated ovarian follicular fluids (FF) contain luteal granulosa cells (LGCs) and other contaminating cell types. Several strategies, such as the antibody binding methods, the flask method, the cell strainer and positive selection of granulosa aggregates after density gradient (DG) centrifugation, were tested as LGC purification methods. Each of these strategies has its own advantages and disadvantages. Positive selection of granulosa aggregates after DG centrifugation is simple, rapid and efficient in terms of LGC recovery. However, it results in a low purity. Here, we aimed to test whether modifying the traditional protocol by collecting the aggregates from the FF, before the DG centrifugation could decrease the percentage of contaminating cells. Materials and Methods: In the present prospective study, 32 FF, from 32 women,were randomly assigned into one of the two purification techniques: positive selection of granulosa aggregates from the FF, after DG centrifugation (DG/ Agg, n=16) or positive selection of granulosa aggregates from the FF, before DG centrifugation (Agg/DG, n=16). At the end of each procedure cell count, vitality, morphology and purity of the cell suspension were evaluated. Results: No significant difference was detected in the total number of GCs between DG/Agg and Agg/DG (P>0.05). However, higher percentage of GCs with normal morphology was detected in Agg/DG compared to DG/Agg (P<0.001). Moreover, lower percentages of white blood cells (P<0.01), red blood cells (P<0.001) and epithelial cells (P<0.01) were identified in Agg/DG compared to DG/Agg. Conclusion: Here we showed that positive selection of granulosa aggregates from the FF prior to DG technique had a higher purity compared to the traditional protocol. Thus, it could be a method of choice to prepare GCs for research purposes in clinical in vitro fertilization settings.
Authors and Affiliations
Georges Raad
Keywords
Related Articles
- EP ID EP669353
- DOI 10.22074/ijfs.2020.5790.
- Views 175
- Downloads 0
Rubella Immunity in Pregnant Iranian Women: A Systematic Review and Meta-Analysis
Rubella infection within the first trimester of pregnancy may lead to adverse pregnancy outcomes. The present study was conducted to evaluate the immunity against rubella among the pregnant Iranian women. The steps of me...
The Frequency of Staphylococcus Aureus Isolated from Endocervix of Infertile Women in Northwest Iran
Background Infertility is one of the major social issues. Due to the asymptomatic cervical infection associated with Staphylococcus aureus (S. aureus), the majority of patients remain undiagnosed. The present study inten...
Human Sperm Quality and Metal Toxicants: Protective Effects of some Flavonoids on Male Reproductive Function
Background Metals can cause male infertility through affection of spermatogenesis and sperm quality. Strong evidences confirm that male infertility in metal-exposed humans is mediated via various mechanisms such as produ...
Varicocele-Induced Infertility in Animal Models
Varicocele is characterized by abnormal tortuosity and dilation of the veins of the pampiniform plexus within the spermatic cord. Although several reports show the mechanisms by which the varicocele exerts its infertilit...
Relationship between Health Literacy and Sexual Function and Sexual Satisfaction in Infertile Couples Referred to The Royan Institute
Background Health science and technology today is a rapidly growing field. Health is a multifaceted concept influ- enced by several factors, and health literacy is essential to deal properly with the current situation. I...