Device and method for cryopreserving animal cells in batch

Abstract

The invention relates to a device and a method for cryopreserving animal cells in batch; the device consists of a cell cryopreservation bag, a cell cryopreservation box and a cell rack; the cell cryopreservation bag is positioned on the cell rack; and the cell rack is positioned inside the cell cryopreservation box. The method for cryopreserving the cells in batch comprises the following steps that: a cell suspension is filled in the cell cryopreservation bag and positioned on the cell rack; the cell rack is put in the cell cryopreservation box; the cell cryopreservation box is thrown into a liquid nitrogen tank or freezing equipment at a temperature of less than 70 below zero for freezing 24 hours; the cell cryopreservation bag is taken out, thrown into liquid nitrogen or the freezing equipment at the temperature of less than 70 below zero and cryopreservated for a long time; when the cells need to be defreezed, the cell cryopreservation bag is pulled out and put in warm water at a temperature of between 38 and 39 DEG C and defreezed; after coagulum is molten into liquid, the liquid is moved to a laminar flow hood; according to a sterile operating procedure, the cell suspension in the cell cryopreservation bag is subpackaged into a centrifuge tube for centrifugation; a supernatant fluid is discarded; and the suspension is operated according to routine cell cryopreservation. The volume of the cryopreserving cell of the cryopreservation bag reaches between 50 and 200 ml; and the whole set of the device is directly thrown into the liquid nitrogen or put in the freezing equipment at the temperature of less than 70 DEG C below zero.

Description

technical field [0001] The invention belongs to the field of biotechnology and biological products, and in particular relates to a device and method for one-step cryopreservation of cells in a large batch in one cryopreservation unit. Background technique [0002] Ultra-low temperature cryopreservation is a technical link that must be experienced in the process of animal cell culture and application, and is widely used in biotechnology research and biological product production industries. At present, people mainly adopt the following methods for cryopreservation of animal cells and their cultures: Digest and disperse the cells with special reagents (such as trypsin, EDTA, etc.), collect them by centrifugation, and then dilute them to a certain density with a special cryoprotectant solution. Or directly disperse the digested cells with a special cryoprotectant solution, divide them into special cryopreservation tubes, and move the cryopreservation tubes step by step into env...

AI Technical Summary

Problems solved by technology

This cryopreservation technology has two obvious defects: 1. For biological product companies that need to culture cells in large quantities, the volume of each cryopreservation unit (1 cryopreservation tube) is only 1-5ml, which can be frozen The amount of cells is extremely limited. After thawing, the cells in each cryopreservation tube are often only enough for a 50-100ml Kerry flask for normal culture. However, almost all large-scale biological products companies use more than 10,000 ml. Some spinner flask culture cells, some even use bioreactors to culture cells, the amount of cells required for a single culture unit may be equivalent to the amount of cells cultured in dozens to hundreds of 100ml Kirschner flasks, therefore, from a It usually takes at least 15 days for the cells in the cryopreservation tube to expand to the amount of cells required to inoculate a spinner bottle. As for the expansion to the amount of cells required to inoculate a bioreactor, it takes longer, so long The waiting time is a great waste for the enterprise; in addition, when the enterprise encounters large-scale pollution, or accidentally obtains the production right of a new product, if it wants to maximize the production capacity of the factory in an instant, it must rely on freezing Expansion of deposited cells is not achievable

2. The cryopreservation process of cells often requires a more complicated procedure, and it needs to be placed at different temperatures for a period of time, which is extremely inconvenient for the operator, and often results in a batch of frozen storage being placed at a certain intermediate temperature for too long. cell scrapping

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