Department of Obstetrics and Gynecology, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.
Reprod Biomed Online. 2011 Feb;22(2):220-7. doi: 10.1016/j.rbmo.2010.10.010. Epub 2010 Oct 31.
Bioluminescence imaging (BLI) has been introduced for studies of ongoing biological processes but has never been applied for ovarian transplantation. Here, BLI was used as a novel approach to trace the survival of ovarian grafts. The ovarian donors were transgenic mice carrying FVB/N-Tg (PolII-luc) as a reporter gene, encoding luciferase to catalyse luciferin which results in visible light emission as bioluminescence. There were three groups of recipients: (i) group A: BALB/c mice without immunosuppressant treatment; (ii) group B: BALB/c mice receiving a cocktail immunosuppressant treatment; and (iii) group C: immunodeficient NOD-SCID mice without immunosuppression. Luciferin BLI was used to follow graft survival, and viable follicle numbers were counted as a measure of success. Bioluminescence intensity fluctuated but was consistent with the end-point counts of viable follicle numbers. Group A showed loss of viable follicles and bioluminescence disappeared as early as day 10 following ovarian engraftment, indicating strong immune rejection. Groups B and C showed graft survival and measurable bioluminescence for up to 30 days. In conclusion, BLI provided non-invasive longitudinal dynamic monitoring of ovarian grafts with excellent sensitivity and spatial resolution. This approach should prove valuable for research on ovarian transplantation.
生物发光成像是一种用于研究正在进行的生物过程的方法，但从未应用于卵巢移植。本研究首次将生物发光成像（BLI）用于追踪卵巢移植物的存活。供体卵巢是携带 FVB/N-Tg（PolII-luc）作为报告基因的转基因小鼠，报告基因编码荧光素酶，可催化荧光素产生可见光发射，即生物发光。实验设置三组受体：（i）组 A：未接受免疫抑制治疗的 BALB/c 小鼠；（ii）组 B：接受鸡尾酒免疫抑制治疗的 BALB/c 小鼠；（iii）组 C：无免疫抑制的免疫缺陷 NOD-SCID 小鼠。使用荧光素 BLI 来跟踪移植物的存活情况，并计数有活力的卵泡数量作为成功的指标。生物发光强度波动，但与有活力的卵泡数量的终点计数一致。组 A 在卵巢移植后第 10 天就表现出有活力的卵泡丢失和生物发光消失，表明强烈的免疫排斥反应。组 B 和组 C 显示移植物存活和可测量的生物发光长达 30 天。总之，BLI 提供了对卵巢移植物的非侵入性纵向动态监测，具有极好的灵敏度和空间分辨率。这种方法对于卵巢移植的研究应该具有重要价值。
