Methodology for processing mastectomy and cryopreservation of breast cancer tissue in a resource- poor setting: A pilot study

在资源匮乏的环境中处理乳房切除术和乳腺癌组织冷冻保存的方法:一项初步研究

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作者:Uzoamaka A Okoli, Michael T Okafor, Kenneth A Agu, Augustine C Ndubuisi, Ifeoma J Nwigwe, Emmanuel O Nna, Okechukwu C Okafor, Francis I Ukekwe, Teresa U Nwagha, Victor C Menkiti, Charles O Eze, Kenechukwu C Onyekwelu, Joy E Ikekpeazu, Chikere A Anusiem, Anthony U Mbah, Chioli P Chijioke, Iroka J Ude

Background

There is scarcity of breast cancer tissues derived from women of African origin available for patient - derived xenograft and organoid models.

Objective

We aim to create a versatile protocol for processing mastectomy and cryopreservation of breast cancer tissue. Methodology: 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% Me2SO) before snap -freezing in liquid Nitrogen vapour and final short-term storage in -80 °C Freezer. The histomorphology, tissue and molecular viability were assessed.

Results

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.

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