Scale-Up Production of Chicken Myoblasts in Microcarrier-Based System

With the rapid development of the cultivated meat industry, large-scale cell culture technology applied to cultivated meat production has received extensive attention. In this study, two microcarriers, Cytodex 1 and 3D TableTrix™, were used for suspension culture of myoblasts in spinner flasks to se...

Full description

Bibliographic Details
Main Author: LIU Yisen, CAI Jiaqi, LI Shilei, LI Yingying, WANG Shouwei, YANG Feng, HU Haijuan, LIU Wenting, LI Yushuang, DONG Shengyan, CHEN Jiaxuan, LIANG Jun
Format: Article
Language:English
Published: China Food Publishing Company 2023-12-01
Series:Shipin Kexue
Subjects:
Online Access:https://www.spkx.net.cn/fileup/1002-6630/PDF/2023-44-24-017.pdf
_version_ 1797365401362890752
author LIU Yisen, CAI Jiaqi, LI Shilei, LI Yingying, WANG Shouwei, YANG Feng, HU Haijuan, LIU Wenting, LI Yushuang, DONG Shengyan, CHEN Jiaxuan, LIANG Jun
author_facet LIU Yisen, CAI Jiaqi, LI Shilei, LI Yingying, WANG Shouwei, YANG Feng, HU Haijuan, LIU Wenting, LI Yushuang, DONG Shengyan, CHEN Jiaxuan, LIANG Jun
author_sort LIU Yisen, CAI Jiaqi, LI Shilei, LI Yingying, WANG Shouwei, YANG Feng, HU Haijuan, LIU Wenting, LI Yushuang, DONG Shengyan, CHEN Jiaxuan, LIANG Jun
collection DOAJ
description With the rapid development of the cultivated meat industry, large-scale cell culture technology applied to cultivated meat production has received extensive attention. In this study, two microcarriers, Cytodex 1 and 3D TableTrix™, were used for suspension culture of myoblasts in spinner flasks to select a more suitable one for large-scale culture of myoblasts, and then the effects of different culture conditions on the large-scale culture of cells were explored. The microscopic results showed that Cytodex 1 was spheres with a smooth surface and no macroporous structure, and 3D TableTrix™ was irregular spheres with a porous surface and large pores. The results of cell culture in spinner flasks showed that the cells grew on 3D TableTrix™ more efficiently, and after 10 days of culture, the cell yield was 8.97 × 105 cells/mL. The optimal conditions for cell attachment were intermittent stirring at 40 r/min for 10 min with intervals of 50 min. The best cell culture efficiency was obtained by using cell inoculation density of 1 × 105 cells/mL, microcarrier density of 2 mg/mL, agitation speed of 40 r/min, and initial culture serum concentration of 20% as well as replacing the medium with a new one containing 10% serum (V/V) after 24 h of culture, and replacing 50% of the medium with a new one with every other day. By bead to bead transfer of the microcarriers, the cells were transferred to a 2 L spinner flask. Finally, 1.07 × 109 cells were harvested with a survival rate of more than 95%. Microcarriers loaded with cells could be cryopreserved and resuscitated, and no significant differences in proliferation status and morphology were observed compared with thawed cells. The optimized microcarrier based culture process will have promising applications in the industrial scale production of cultivated meat.
first_indexed 2024-03-08T16:48:22Z
format Article
id doaj.art-c3dc2b4481e5436b867c8ee8bfebc58c
institution Directory Open Access Journal
issn 1002-6630
language English
last_indexed 2024-03-08T16:48:22Z
publishDate 2023-12-01
publisher China Food Publishing Company
record_format Article
series Shipin Kexue
spelling doaj.art-c3dc2b4481e5436b867c8ee8bfebc58c2024-01-05T07:16:26ZengChina Food Publishing CompanyShipin Kexue1002-66302023-12-01442412713710.7506/spkx1002-6630-20230330-310Scale-Up Production of Chicken Myoblasts in Microcarrier-Based SystemLIU Yisen, CAI Jiaqi, LI Shilei, LI Yingying, WANG Shouwei, YANG Feng, HU Haijuan, LIU Wenting, LI Yushuang, DONG Shengyan, CHEN Jiaxuan, LIANG Jun0(1. State Key Laboratory of Food Nutrition Safety, Tianjin University of Science & Technology, Tianjin 300457, China; 2. China Meat Research Center, Beijing Key Laboratory of Meat Processing Technology, Beijing Academy of Food Sciences, Beijing 100068, China)With the rapid development of the cultivated meat industry, large-scale cell culture technology applied to cultivated meat production has received extensive attention. In this study, two microcarriers, Cytodex 1 and 3D TableTrix™, were used for suspension culture of myoblasts in spinner flasks to select a more suitable one for large-scale culture of myoblasts, and then the effects of different culture conditions on the large-scale culture of cells were explored. The microscopic results showed that Cytodex 1 was spheres with a smooth surface and no macroporous structure, and 3D TableTrix™ was irregular spheres with a porous surface and large pores. The results of cell culture in spinner flasks showed that the cells grew on 3D TableTrix™ more efficiently, and after 10 days of culture, the cell yield was 8.97 × 105 cells/mL. The optimal conditions for cell attachment were intermittent stirring at 40 r/min for 10 min with intervals of 50 min. The best cell culture efficiency was obtained by using cell inoculation density of 1 × 105 cells/mL, microcarrier density of 2 mg/mL, agitation speed of 40 r/min, and initial culture serum concentration of 20% as well as replacing the medium with a new one containing 10% serum (V/V) after 24 h of culture, and replacing 50% of the medium with a new one with every other day. By bead to bead transfer of the microcarriers, the cells were transferred to a 2 L spinner flask. Finally, 1.07 × 109 cells were harvested with a survival rate of more than 95%. Microcarriers loaded with cells could be cryopreserved and resuscitated, and no significant differences in proliferation status and morphology were observed compared with thawed cells. The optimized microcarrier based culture process will have promising applications in the industrial scale production of cultivated meat.https://www.spkx.net.cn/fileup/1002-6630/PDF/2023-44-24-017.pdfmyoblast; cultivated meat; microcarriers; spinner flask; optimized culture system
spellingShingle LIU Yisen, CAI Jiaqi, LI Shilei, LI Yingying, WANG Shouwei, YANG Feng, HU Haijuan, LIU Wenting, LI Yushuang, DONG Shengyan, CHEN Jiaxuan, LIANG Jun
Scale-Up Production of Chicken Myoblasts in Microcarrier-Based System
Shipin Kexue
myoblast; cultivated meat; microcarriers; spinner flask; optimized culture system
title Scale-Up Production of Chicken Myoblasts in Microcarrier-Based System
title_full Scale-Up Production of Chicken Myoblasts in Microcarrier-Based System
title_fullStr Scale-Up Production of Chicken Myoblasts in Microcarrier-Based System
title_full_unstemmed Scale-Up Production of Chicken Myoblasts in Microcarrier-Based System
title_short Scale-Up Production of Chicken Myoblasts in Microcarrier-Based System
title_sort scale up production of chicken myoblasts in microcarrier based system
topic myoblast; cultivated meat; microcarriers; spinner flask; optimized culture system
url https://www.spkx.net.cn/fileup/1002-6630/PDF/2023-44-24-017.pdf
work_keys_str_mv AT liuyisencaijiaqilishileiliyingyingwangshouweiyangfenghuhaijuanliuwentingliyushuangdongshengyanchenjiaxuanliangjun scaleupproductionofchickenmyoblastsinmicrocarrierbasedsystem