Vitrification-based Cryopreservation: An Ultra-rapid Cooling Technique to Preserve Biological Specimens at Very Low Temperatures for Long-term Storage

Abstract

Source: Lu, J. H. et al. Establishment of Gastric Cancer Patient-derived Xenograft Models and Primary Cell Lines. J. Vis. Exp. (2019)

In this video, we demonstrate an ultra-rapid cryo vitrification method for preserving biological samples. The method entails the use of a combination of cryoprotectants to protect the biological tissue from freezing damage by preventing intracellular ice crystal formation.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Tissue Cryopreservation

NOTE: This part primarily references the methods for the Live Tissue Kit Cryo Kit. The main kits and equipment are listed in the Table of Materials.

1. Euthanize the mice with an IRB-approved method when the tumor is greater than 10 mm in diameter.
2. Using sterile forceps and scissors, slowly isolate the tumor from the mice.
3. Wash the tumor tissues with DPBS in a ....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Live Tissue Kit Cryo Kit	Celliver Biotechnology, Shanghai, China	LT2601
Tissue-processed molds and auxiliary blades	Celliver Biotechnology, Shanghai, China	LT2603
DPBS	Basalmedia Technology, Shanghai, China	L40601