Saadeldin Islam M, Elsayed Asmaa, Kim Su Jin, Moon Joon Hu, Lee Byeong Chun
Indian J Exp Biol. 2015 Feb;53(2):75-81.
The bidirectional communication between oocytes and granulosa cells are mediated by several factors via a local feedback loop(s). The current model was carried out to study the spatial mutual interaction of porcine denuded oocytes and granulosa cells either in direct contact (juxtacrine) or paracrine co-culture using transwell system. Transwell 0.4 μm polyester membrane inserts were used to permit oocytes-granulosa cells paracrine communication with a distance of 2 mm between them in co-culture. Oocytes were cultured with granulosa cells in a defined basic maturation medium for 44 h. In results, oocyte secreted factors (OSFs; GDF9 and BMP15) temporal expression showed progressive decrement by the end of culture in case of direct contact with granulosa cells while it was increased progressively in the paracrine co-culture groups. However, oocytes that were cultured in direct contact showed a significant increase in blastocyst development after parthenogenetic activation than the paracrine co-cultured ones (20% vs. 11.5%, respectively). By the end of culture, granulosa cell count in direct contact showed a significant decrease than the indirect co-culture group (1.2 x 105 cell/mL vs. 2.1 x 10(5) cell/mL, respectively). Steroids (P4 and E2) and steriodogenesis enzymes mRNA levels showed significant temporal alterations either after 22 h and 44 h of IVM in both juxtacrine and paracrine co-culture systems (P ≤ 0.05). CX43 was much more highly expressed in the granulosa of the direct contact group than the indirect co-culture group. These results indicate the difference in mutual communication between oocytes and granulosa cells that were cocultured either in direct contact (juxtacrine) or with a short distance (paracrine) and propose a new paradigm to study different ovarian follicular cells interaction.
卵母细胞与颗粒细胞之间的双向通讯是由多种因素通过局部反馈回路介导的。当前模型旨在研究猪裸卵母细胞与颗粒细胞在直接接触（旁分泌）或使用Transwell系统进行旁分泌共培养时的空间相互作用。使用Transwell 0.4μm聚酯膜插入物，使卵母细胞与颗粒细胞在共培养时相距2mm进行旁分泌通讯。卵母细胞与颗粒细胞在确定的基础成熟培养基中培养44小时。结果显示，与颗粒细胞直接接触时，卵母细胞分泌因子（OSFs；GDF9和BMP15）的时间表达在培养结束时逐渐下降，而在旁分泌共培养组中则逐渐增加。然而，直接接触培养的卵母细胞在孤雌激活后囊胚发育的显著增加高于旁分泌共培养的卵母细胞（分别为20%和11.5%）。培养结束时，直接接触组的颗粒细胞计数显著低于间接共培养组（分别为1.2×10⁵细胞/mL和2.1×10⁵细胞/mL）。在旁分泌和旁分泌共培养系统中，IVM 22小时和44小时后，类固醇（P4和E2）和类固醇生成酶mRNA水平均显示出显著的时间变化（P≤0.05）。CX43在直接接触组颗粒细胞中的表达远高于间接共培养组。这些结果表明，卵母细胞与颗粒细胞在直接接触（旁分泌）或短距离（旁分泌）共培养时相互通讯存在差异，并提出了一种研究不同卵巢卵泡细胞相互作用的新范式。
