Department of Medical Biochemistry and Molecular Biology, College of Medicine, University of Nigeria Enugu Campus, Nigeria; Department of Pharmacology and Therapeutics, College of Medicine, University of Nigeria Enugu Campus, Nigeria.
Department of Pharmacology and Therapeutics, College of Medicine, University of Nigeria Enugu Campus, Nigeria.
Cryobiology. 2020 Dec;97:179-184. doi: 10.1016/j.cryobiol.2020.05.006. Epub 2020 Jun 18.
There is scarcity of breast cancer tissues derived from women of African origin available for patient - derived xenograft and organoid models.
We aim to create a versatile protocol for processing mastectomy and cryopreservation of breast cancer tissue.
An immediate collection of breast cancer tissue from mastectomy was bathed in 4 °C HBSS and immediately transferred to 4 °C RPMI1640 containing HEPES, 10% FBS, Streptomycin and Penicillin. Tissues were processed over ice yielding nine samples of cold ischemic time (20-45 min) stored at 3 min interval. Cut samples were transferred into cryovials containing 4 °C cryoprotectant agent (90% FBS +10% MeSO) before snap -freezing in liquid Nitrogen vapour and final short-term storage in -80 °C Freezer. The histomorphology, tissue and molecular viability were assessed.
The cold ischemic times had no detrimental effect to the nine samples despite being processed in a resource poor setting, hence providing a reproducible and reliable protocol.
用于患者来源的异种移植物和类器官模型的源自非洲裔女性的乳腺癌组织稀缺。
我们旨在创建一种用于处理乳腺癌组织的乳房切除术和冷冻保存的多功能方案。
从乳房切除术中立即采集乳腺癌组织，置于 4°C HBSS 中，并立即转移至含有 HEPES、10% FBS、链霉素和青霉素的 4°C RPMI1640 中。组织在冰上进行处理，产生 9 个冷缺血时间（20-45 分钟）的样本，每隔 3 分钟存储一次。切下的样本转移到含有 4°C 冷冻保护剂（90% FBS+10% MeSO）的冷冻管中，然后在液氮蒸气中快速冷冻，并在-80°C 冰箱中进行短期储存。评估了组织学形态、组织和分子活力。
尽管在资源匮乏的环境中进行处理，但 9 个样本的冷缺血时间没有产生不利影响，因此提供了一种可重复和可靠的方案。
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