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Serum-free cell freezing medium

A cryopreservation liquid and serum-free technology, which is applied in the field of cell culture, can solve the problems of increased animal pathogen contamination, poor cryopreservation effect, and failure of clinical infusion, etc., and achieves low cost, good application prospects, and good adherent growth Effect

Inactive Publication Date: 2018-08-28
成都菱祐生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows for easy storage and transporting without harmful substances like water during freezing procedures while still maintaining their effectiveness at delivering therapeutic agents safely into patients' bloodstream.

Problems solved by technology

This patented technical problem addressed in this patent relates to improving the safety and efficacy of culturing and recovering living tissues from cellular suspensions during cryotherapy procedures while reducing harmful effects associated with these treatments such as exposure to chemical agents like formaldehyde, ethylenegleneimines, histidase enzymes, etc., and without requiring complicated methods involving expensive materials containing specific components.

Method used

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  • Serum-free cell freezing medium
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Examples

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Effect test

Embodiment 1

[0025] Cryopreservation of mouse compact bone mesenchymal stem cells (CBMSCs):

[0026] Mouse compact bone mesenchymal stem cells (CBMSCs) were isolated and cultured using the tissue block attachment method. The mouse femur and humerus were cut into pieces and attached to a 10 cm culture dish, and DME / F12 medium was added for culture. It was digested regularly and the cells were collected for cryopreservation.

[0027] According to the volume ratio, mix 90-99% CD medium with 1-10% DMSO to prepare cell cryopreservation solution, which is prepared and used immediately.

[0028] The above-mentioned expanded mesenchymal stem cells were divided into 5×10 6 Each was resuspended in 1.5 parts of serum-free cryopreservation solution, and then the cell suspension was added to a cryopreservation tube, which was placed in a programmed cooling box at normal temperature, and then the programmed cooling box was placed at -80°C ultra-low temperature Refrigerate overnight and transfer to a l...

Embodiment 2

[0031] Cryopreservation of mouse breast cancer (4T1) cells:

[0032] Take 1 dish of 4T1 cells in the logarithmic growth phase within 40 passages, discard the supernatant, add sulfate buffer to wash once, add trypsin to digest, and then add serum-containing medium to stop digestion, and put the The cell suspension was added to the centrifuge tube, and the supernatant was discarded after centrifugation.

[0033] According to the volume ratio, mix 90-99% CD medium with 1-10% DMSO to prepare cell cryopreservation solution, which is prepared and used immediately.

[0034] Take 5×10 6 2 cells were added to 1.5 parts of serum-free freezing solution for cell resuspension, then the cell suspension was added to the cryopreservation tube, the cryopreservation tube was placed in the programmed cooling box at normal temperature, and then the programmed cooling box was placed Store in a -80°C ultra-low temperature refrigerator overnight, and transfer to a liquid nitrogen tank freezer for ...

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Abstract

The invention discloses a serum-free cell freezing medium and belongs to the field of cell culture. The serum-free freezing medium disclosed by the invention is prepared from: 90-99% of CD media (chemically defined media) and 1-10% of cell culture level dimethyl sulfoxide (DMSO). Without animal-source serum, the freezing medium does not introduce heterologous protein and thus reduces the possibility of animal disease-source contamination; after recovery, the activity of a frozen cell can reach 80-90% while the cell phenotype barely changes, thus the physiological functions and the biological functions of the recovered cell are maintained, and the cell can be further applied to culture or clinical direct feedback. The serum-free freezing medium is safe and effective, the preparation methodis simple, the cost is controllable, and the clinical application prospect is good.

Description

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Claims

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Application Information

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Owner 成都菱祐生物科技有限公司
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