Hoang Nghia Son^1,2, Ho Nguyen Quynh Chi^1,2 and Le Thanh Long^1,2,*

This study aimed to assess morphological changes of porcine granulosa cells (pGCs) following in vitro long-term culture. The pGCs from passage 5 showed a dramatic enlargement of cytoplasm (14422.600±1300.704 µm²) which was 8.23-fold higher than pGCs from primary culture (1752.100±102.244 µm²). Nuclear areas were increased from passage 0 (164.990±3.461 µm²) to passage 4 (399.514±15.110 µm²) and passage 5 (416.326±32.683 µm²), respectively. The nucleocytoplasmic ratio of pGCs from passage 0 was 0.089±0.002 which was 2-fold higher than passage 4 (0.045±0.002) and 3-fold higher than passage 5 (0.029±0.002). The diameter of microfilament bundles of pGCs was increased from passage 0 (1.171±0.031 µm) to passage 4 (1.550±0.056 µm) and passage 5 (1.579±0.053 µm), respectively. The weak expression of beta-galactosidase was observed in the several pGCs from passage 2, while pGCs from passage 4 showed the strong activity of beta-galactosidase. The flow cytometry analysis demonstrated that the ratio of apoptotic pGCs was increased through in vitro expansion. These results revealed that pGCs exposed replicative senescent characteristics including the increase of nuclear and cell size, as well as the increase of diameter of actin filament bundles in cytoplasm, and the reduction of nucleocytoplasmic ratio.

To Cite This Article: Son HN, Chi HNQ and Long LT, 2020. Morphological changes of porcine granulosa cells during in vitro expansion. Pak Vet J, 40(2): 229-233. http://dx.doi.org/10.29261/pakvetj/2020.010