Presentation Information
[2Ip03]Toward cultivated meat production: Biocompatibility analysis of propylene glycol-based cryopreservation solution solely prepared by food and food additives
*Takuya Sugiura1,3, Kazumasa Nodake1, Akiko Takasuga1, Miki Mizukami2, Koji Nakade1, Tetsusei Kurashiki3, Michiya Matsusaki3 (1. ITOHAM YONEKYU HOLDINGS INC., 2. TOPPAN Holdings Inc., 3. Osaka University)
Keywords:
Cultured meat,Cryopreservation solution,Propylene glycol,the Food Sanitation Act,the Standards for Use of Food Additives
【Purpose】We have customized a commercially available cell cryopreservation solution (STEM-CELLBANKER DMSO Free GMP grade, ZENOGEN PHARMA) to consist only of food and food additives. However, this cryopreservation solution contains propylene glycol (PG) (10% [v/v]) at a higher concentration than the standard for the use of food additives (0.60% [w/w], the upper limit for use in general foods), so we aimed to specify the process in which the PG content in the cells is less than 0.60% [w/w]. In addition, we investigated whether or not this cryopreservation solution could sufficiently maintain the function of the source cells of cultured meat.【Methods】
Bovine muscle satellite cells (bSCs) and adipose tissue-derived stem cells (bADSCs) were cryopreserved in the custom-made cryopreservation solution. The PG content in the cells was determined after the cryopreserved cells were washed for seeding and cultured for expansion. In addition, the function of the cryopreserved cells after 6 months of storage was evaluated by cell viability, proliferation and differentiation potential.【Results】The PG in the cryopreserved cells decreased quickly by a general cell washing method and disappeared during expansion culture; washing with 45 times the volume of medium reduced the PG content in the recovered cells to less than half the regulation value. Thus, the PG concentration in the cells could be maintained well below the regulatory level after recovery from cryopreservation, indicating that the risk of food hazards due to PG could be sufficiently reduced. Furthermore, cells stored in this cryopreservation solution for 6 months maintained sufficient cell viability, proliferative and differentiation potential.
Bovine muscle satellite cells (bSCs) and adipose tissue-derived stem cells (bADSCs) were cryopreserved in the custom-made cryopreservation solution. The PG content in the cells was determined after the cryopreserved cells were washed for seeding and cultured for expansion. In addition, the function of the cryopreserved cells after 6 months of storage was evaluated by cell viability, proliferation and differentiation potential.【Results】The PG in the cryopreserved cells decreased quickly by a general cell washing method and disappeared during expansion culture; washing with 45 times the volume of medium reduced the PG content in the recovered cells to less than half the regulation value. Thus, the PG concentration in the cells could be maintained well below the regulatory level after recovery from cryopreservation, indicating that the risk of food hazards due to PG could be sufficiently reduced. Furthermore, cells stored in this cryopreservation solution for 6 months maintained sufficient cell viability, proliferative and differentiation potential.
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