Two techniques for the isolation and purification of ovarian GCs (granulosa cells) were compared and analysed, with the aim of providing an efficient and simple solution for obtaining GCs from patients with DOR(diminished ovarian reserve). 160 follicular fluids from DOR patients undergoing in vitro fertilization or intracytoplasmic sperm injection were collected and randomly divided into P/DG group(modified traditional density gradient centrifugation group) and W/Agg group(granulosa cell aggregation washing group), and the extracted GCs were identified by purity, evaluated for proliferation, and analyzed by pathology, morphology and statistics. The results indicate that GCs extracted from both groups predominantly aggregate into clusters, with cells interacting via filamentous protrusions. Lipid droplets are visible under microscopy, and adherent cell growth begins within 24 ~ 48 h. Flow cytometry analysis reveals that the W/Agg group exhibits slightly lower suspension purity but significantly reduces total processing time while increasing cell yield and viability (P < 0.05). HE staining and FSHR immunofluorescence confirm that GCs in both groups maintain intact morphology, displaying a pike- or star-shaped appearance, with the W/Agg group showing higher cell density and aggregation (P <0.05). CCK-8 dynamic monitoring demonstrates enhanced proliferation in the W/Agg group by day 6 (P < 0.05), with a higher peak activity (P < 0.05). The W/Agg technique proves superior in operational efficiency, cell yield, and viability. Although initial purity is marginally lower, subsequent cultures achieve higher cell density and purity. These findings provide a robust scientific foundation for optimizing GC isolation, purification, and in vitro culture protocols, particularly for patients with DOR.