模拟网膜卵巢癌细胞定植:一种在小鼠网膜外植体中建立荧光标记卵巢癌细胞银斑定植的离体方法

大网膜是腹膜腔内富含脂肪的组织，由脂肪库组成，为转移性癌细胞定植并形成继发性肿瘤提供合适的微环境。要研究这种离体定植，首先将可渗透的膜细胞培养插入物放置在培养板内。

插入膜的内部区域涂上组织粘合剂。将小鼠大网膜外植体放在粘合剂上以固定组织。轻轻地将荧光标记的卵巢癌细胞悬浮液接种在大网膜顶部。

接下来，用合适的培养基填充孔的侧面以补充细胞生长并孵育所需的时间。大网膜组织包含称为乳白色斑点的特殊结构，乳状斑点是围绕血管网络的免疫细胞簇。这些免疫细胞会分泌某些化学因子来吸引癌细胞并促进其着床。定植的癌细胞增殖，模拟转移条件。

最后，丢弃用过的介质。使用荧光显微镜观察培养物以评估大网膜组织内的荧光癌细胞集落。

培养和制备荧光标记细胞，并以每毫升 200 万个细胞的浓度重悬。将大约 6 微升组织粘合剂涂在培养插入物的膜上，并使其风干。然后，用无菌水清洗膜两次，去除任何多余的粘合剂，然后在层流罩下风干膜。

小心地切除大网膜，并使用无菌镊子将其附着在涂有粘合剂的膜上。让组织粘附在膜上一分钟后，在每个培养物插入物中的每个网膜顶部加入500微升细胞悬液。

然后，用 2.5 毫升 DME/F-12 培养基填充 transwell 室周围的区域。在5%CO2环境中将大网膜与细胞悬液在37摄氏度下孵育6小时。小心地取出并用约 10 毫升 PBS 清洗大网膜。最后，使用适当的荧光成像系统可视化荧光癌细胞病灶。

• Omentum – An adipose-rich tissue within the peritoneal cavity.
• Omental Tissue – Tissue providing a microenvironment for cancer cells.
• Ovarian Cancer – Cancer that seeds and proliferates on the omentum.
• Milky Spots – Structures that attract and support cancer cell implantation.
• Omental Metastasis – Secondary tumor colonization within the omental tissue.

• Omentum is a fat-rich depot providing a home for cancer cells (e.g., omental tissue).
• The omental tissue contains specialized structures known as milky spots (e.g., theory).
• Milky spots secrete chemicals, attracting and supporting cancer cells (e.g., processes).
• The seeded cancer cells proliferate, simulating metastatic conditions (e.g., experiment).

• What is the role of omentum and milky spots in ovarian cancer metastasis?
• What causes proliferation of cancer cells within the omental tissue?
• How do milky spots support cancer cell implantation?

• This research can contribute to new treatment strategies for ovarian cancer (e.g., technology).
• The findings can impact healthcare by improving understanding of metastasis (e.g., healthcare).
• The research could lead to improved quality of life for patients with ovarian cancer (e.g., sustainability).
• The research paves the way for advancement in the treatment of metastatic cancers (e.g., innovations).