A novel ovarian xenografting model to characterize the impact of chemotherapy agents on human primordial follicle reserve
A novel ovarian xenografting model to characterize the impact of chemotherapy agents on human primordial follicle reserve
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DOI：
10.1158/0008-5472.can-07-2042
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发表时间：
2007-11-01
期刊：
影响因子：
11.2
通讯作者：
Oktay, Kutluk
中科院分区：
文献类型：
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作者：
Oktem, Ozgur;Oktay, Kutluk
许多化疗药物，尤其是烷化剂家族的化疗药物，会改变绝经前女性的生育能力。然而，在临床环境中量化和表征癌症药物对卵巢储备的影响实际上是不可能的。因此，我们的具体目标是开发异种移植模型来表征化疗药物对人类卵巢的体内影响。将来自 24 周龄流产者的卵巢碎片进行皮下异种移植。严重联合免疫缺陷小鼠 (n = 52)。动物接受单剂量 200 mg/kg 的环磷酰胺或载体。环磷酰胺注射后 12 至 72 小时，从对照小鼠和治疗小鼠中回收移植物，并连续切片进行原始卵泡计数。通过末端核苷酸转移酶介导的切口末端标记（TUNEL）测定来评估细胞凋亡。通过共聚焦显微镜进行血小板/内皮细胞粘附分子 I (PECAM-1) 染色以及活体内荧光素缀合的凝集素和伊文思蓝标记来评估微脉管系统。虽然治疗后12小时原始卵泡密度减少12%（P>0.05），但卵泡损失在24小时时显着增加（53%，P<0.01），并在48小时时达到峰值（93%，P<0.0001）。 TUNEL 染色在 12 小时达到峰值，早于卵泡数量的减少，此后逐渐减少。异种移植血管化模式与非异种移植组织相似，表明体内药物输送适当。在我们的人类卵巢异种移植模型中，环磷酰胺对原始卵泡储备的影响与该药物的临床性腺毒性一致。人类卵巢异种移植是一种很有前途的模型，可以表征当前和新兴癌症药物的性腺毒性作用，而无需进行漫长的临床研究。
Many chemotherapeutic agents, especially of the alkylating family, alter fertility in premenopausal females. However, it is not practically possible to quantify and characterize the impact of cancer drugs on ovarian reserve in a clinical setting. Thus, our specific aim was to develop a xenograft model to characterize the in vivo impact of chemotherapy agents on human ovary. Ovarian pieces from 24 weeks old abortuses were xenografted s.c. to severe combined inummodeficient mice (n = 52). Animals received either a single dose of 200 mg/kg of cyclophosphamide or the vehicle. Grafts were recovered from the control and treated mice 12 to 72 h after the cyclophosphamide injection and serially sectioned for primordial follicle counts. Apoptosis was assessed with terminal nucleotidyl transferase-mediated nick end labeling (TUNEL) assay. Platelet/endothelial cell adhesion molecule I (PECAM-1) staining, as well as intra-vital fluorescein-conjugated lectin and Evans blue labeling were done to assess microvasculature by confocal microscopy. Although there was 12% reduction in primordial follicle density by 12 h following treatment (P > 0.05), the follicle loss increased significantly at 24 h (53%, P < 0.01) and peaked at 48 h (93%, P < 0.0001). TUNEL staining peaked at 12 h, earlier than the diminishment in follicle numbers, and decreased thereafter. Xenograft vascularization pattern was similar to non-xenografted tissue, indicating appropriate in vivo drug delivery. The impact of cyclophosphamide on primordial follicle reserve in our human ovarian xenograft model is consistent with the clinical gonadotoxicity of this drug. Human ovarian xenografting is a promising model to characterize the gonadotoxic effects of current and emerging cancer drugs without a need for lengthy clinical studies.