The production of tissue-engineered skeletal muscles for cell-based therapy and cultivated meat production re-quires a large number of myogenic cells capable of differentiating into myotubes.A suspension culture with ...The production of tissue-engineered skeletal muscles for cell-based therapy and cultivated meat production re-quires a large number of myogenic cells capable of differentiating into myotubes.A suspension culture with a stirred bioreactor is a system that is feasibly scalable,but scaling up the production of myogenic cells while maintaining differentiation capacity requires an efficient culture technique.This remains challenging in sus-pension culture.On the other hand,conventional meat processing generates substantial by-products,including meat trimmings containing effective substances for myogenic cell cultures.Efficiently utilizing these by-products can reduce environmental impact and disposal costs.Here,we describe an expansion culture method for bovine myogenic cells(BMCs)using Dulbecco’s Modified Eagle’s medium(DMEM)supplemented with muscle extracts.During the 6-day planar culture,the proliferation rate using the culture medium supplemented with muscle extracts(CMME)and 10%fetal bovine serum(FBS)was 18±3.3,three times higher than that achieved using DMEM with 10%FBS.Furthermore,a combination of CMME supplemented with 10%FBS and an iMatrix-511-coated dish could maintain differentiation capacity through passaging culture.Finally,we demonstrated a sus-pension culture with microcarriers using CMME with 10%FBS.Through 3 passages,the cumulative proliferation rate was 191.7±35.1,four times higher than that of produced DMEM with 10%FBS.Using CMME,the cells maintained differentiation capacity in comparison to that using DMEM.This approach will contribute to both the reduction of wasted by-products from conventional meat production and the development of a suspension cul-ture system for maintaining high-quality myogenic cells.展开更多
The electrical conductivity(EC)of extracted muscle fluid has been extensively used to evaluate meat freshness and shelf life in the field of food sanitation for decades.The opposite of freshness is the corruption that...The electrical conductivity(EC)of extracted muscle fluid has been extensively used to evaluate meat freshness and shelf life in the field of food sanitation for decades.The opposite of freshness is the corruption that increases with time.Based on the freshness/corruption principle,we investigated the relationship between long postmortem intervals(PMIs)and EC in cadaver skeletal muscle.EC values of extracted fluid from rat muscles were measured at different PMIs for 10 days.The results indicate that there was a significant correlation between PMI and EC,and the data fit well to the cubic polynomial regression equation y=‑0.01x3+0.264x2‑13.657x+1769.148(R2=0.925).In addition,the EC of different dilutions of these muscle extracts showed strict quadratic correlation(R2=1)with the dilution ratios,suggesting that EC can be measured with very small quantities of muscle sample.Our study suggests that determination of the EC of cadaver skeletal muscle extracts may be a useful method for estimating long PMIs.展开更多
基金supported by the JST-Mirai Program Grant Number JPMJMI20C1,Japan.
文摘The production of tissue-engineered skeletal muscles for cell-based therapy and cultivated meat production re-quires a large number of myogenic cells capable of differentiating into myotubes.A suspension culture with a stirred bioreactor is a system that is feasibly scalable,but scaling up the production of myogenic cells while maintaining differentiation capacity requires an efficient culture technique.This remains challenging in sus-pension culture.On the other hand,conventional meat processing generates substantial by-products,including meat trimmings containing effective substances for myogenic cell cultures.Efficiently utilizing these by-products can reduce environmental impact and disposal costs.Here,we describe an expansion culture method for bovine myogenic cells(BMCs)using Dulbecco’s Modified Eagle’s medium(DMEM)supplemented with muscle extracts.During the 6-day planar culture,the proliferation rate using the culture medium supplemented with muscle extracts(CMME)and 10%fetal bovine serum(FBS)was 18±3.3,three times higher than that achieved using DMEM with 10%FBS.Furthermore,a combination of CMME supplemented with 10%FBS and an iMatrix-511-coated dish could maintain differentiation capacity through passaging culture.Finally,we demonstrated a sus-pension culture with microcarriers using CMME with 10%FBS.Through 3 passages,the cumulative proliferation rate was 191.7±35.1,four times higher than that of produced DMEM with 10%FBS.Using CMME,the cells maintained differentiation capacity in comparison to that using DMEM.This approach will contribute to both the reduction of wasted by-products from conventional meat production and the development of a suspension cul-ture system for maintaining high-quality myogenic cells.
文摘The electrical conductivity(EC)of extracted muscle fluid has been extensively used to evaluate meat freshness and shelf life in the field of food sanitation for decades.The opposite of freshness is the corruption that increases with time.Based on the freshness/corruption principle,we investigated the relationship between long postmortem intervals(PMIs)and EC in cadaver skeletal muscle.EC values of extracted fluid from rat muscles were measured at different PMIs for 10 days.The results indicate that there was a significant correlation between PMI and EC,and the data fit well to the cubic polynomial regression equation y=‑0.01x3+0.264x2‑13.657x+1769.148(R2=0.925).In addition,the EC of different dilutions of these muscle extracts showed strict quadratic correlation(R2=1)with the dilution ratios,suggesting that EC can be measured with very small quantities of muscle sample.Our study suggests that determination of the EC of cadaver skeletal muscle extracts may be a useful method for estimating long PMIs.
