Journal ArticleDOI
Bioprocess development for mass production of size-controlled human pluripotent stem cell aggregates in stirred suspension bioreactor.
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TLDR
In conclusion, suspension culture protocols of hPSCs could be used to mass produce homogenous and pluripotent undifferentiated cells by identification and optimization of key bioprocess parameters that are complemented by a simple and rapid scale-up platform.Abstract:
Current protocols for the scalable suspension culture of human pluripotent stem cells (hPSCs) are limited by multiple biological and technical challenges that need to be addressed before their use in clinical trials. To overcome these challenges, we have developed a novel bioprocess platform for large-scale expansion of human embryonic and induced pluripotent stem cell lines as three-dimensional size-controlled aggregates. This novel bioprocess utilizes the stepwise optimization of both static and dynamic suspension culture conditions. After screening eight xeno-free media in static suspension culture and optimizing single-cell passaging in dynamic conditions, the scale-up from a static to a dynamic suspension culture in the stirred bioreactor resulted in a two- to threefold improvement in expansion rates, as measured by cell counts and metabolic activity. We successfully produced size-specific aggregates through optimization of bioreactor hydrodynamic conditions by using combinations of different agitati...read more
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Scientific, sustainability and regulatory challenges of cultured meat
Mark J. Post,Shulamit Levenberg,David L. Kaplan,Nicholas Genovese,Jianan Fu,Christopher Bryant,Nicole Negowetti,Karin E Verzijden,Panagiota Moutsatsou +8 more
TL;DR: The scientific and social challenges in transforming cultured meat into a viable commercial option are reviewed, covering aspects from cell selection and medium optimization to biomaterials, tissue engineering, regulation and consumer acceptance.
Journal ArticleDOI
Scalable expansion of human induced pluripotent stem cells in the defined xeno-free E8 medium under adherent and suspension culture conditions.
Ying Wang,Bin Kuan Chou,Sarah N. Dowey,Chaoxia He,Chaoxia He,Sharon Gerecht,Linzhao Cheng,Linzhao Cheng +7 more
TL;DR: This system is the first to enable an efficient scale-up bioprocess in completely xeno-free condition for the expansion and cryopreservation of hiPSCs with the quantity and quality compliant for clinical applications.
Journal ArticleDOI
A 3D Sphere Culture System Containing Functional Polymers for Large-Scale Human Pluripotent Stem Cell Production
Tomomi Otsuji,Jiang Bin,Azumi Yoshimura,Misayo Tomura,Daiki Tateyama,Itsunari Minami,Yoshihiro Yoshikawa,Kazuhiro Aiba,John E. Heuser,Taito Nishino,Kouichi Hasegawa,Kouichi Hasegawa,Norio Nakatsuji +12 more
TL;DR: A simple 3D sphere culture system that incorporates mechanical passaging and functional polymers is reported that resolves major problems associated with suspension culture methods and dynamic stirring systems and may be optimal for applications involving large-scale hPSC production.
Journal ArticleDOI
Progress and challenges in large-scale expansion of human pluripotent stem cells
TL;DR: Recent progress in the large-scale expansion of hPSCs with particular focus on suspension culture, which represents a universal strategy for controlled mass cell production, and bioreactor-based approaches, including technical aspects ofBioreactor technologies and operation modes are reviewed.
Journal ArticleDOI
Advances in cell culture: anchorage dependence
TL;DR: Critical issues common to all cell culture systems for adherent cells as well as specifics for different types of stem cells in view of small- and large-scale cell expansion and production processes are given.
References
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Journal ArticleDOI
A ROCK inhibitor permits survival of dissociated human embryonic stem cells
Kiichi Watanabe,Morio Ueno,Daisuke Kamiya,Ayaka Nishiyama,Michiru Matsumura,Takafumi Wataya,Jun Takahashi,Satomi Nishikawa,Shin-Ichi Nishikawa,Keiko Muguruma,Yoshiki Sasai +10 more
TL;DR: Application of a selective Rho-associated kinase (ROCK) inhibitor, Y-27632, to hES cells markedly diminishes dissociation-induced apoptosis, increases cloning efficiency and facilitates subcloning after gene transfer, and enables SFEB-cultured hES Cells to survive and differentiate into Bf1+ cortical and basal telencephalic progenitors.
Journal ArticleDOI
Chemically defined conditions for human iPSC derivation and culture
Guokai Chen,Daniel R. Gulbranson,Daniel R. Gulbranson,Zhonggang Hou,Zhonggang Hou,Jennifer M. Bolin,Victor Ruotti,Mitchell D. Probasco,Kimberly Smuga-Otto,Sara E. Howden,Sara E. Howden,Nicole R. Diol,Nicole R. Diol,Nicholas E. Propson,Nicholas E. Propson,Ryan J. Wagner,Ryan J. Wagner,Garrett O. Lee,Garrett O. Lee,Jessica Antosiewicz-Bourget,Joyce M.C. Teng,James A. Thomson +21 more
TL;DR: A cell culture system in which all protein reagents for liquid media, attachment surfaces and splitting are chemically defined is formed, and improved derivation efficiencies of vector-free human iPSCs with an episomal approach are demonstrated.
Journal ArticleDOI
Biodegradable poly (e-caprolactone) nanoparticles for tumor-targeted delivery of tamoxifen
TL;DR: Results of the study suggest that nanoparticle formulations of selective ER modulators, like tamoxifen, would provide increased therapeutic benefit by delivering the drug in the vicinity of the ER.
Journal ArticleDOI
Synthetic peptide-acrylate surfaces for long-term self-renewal and cardiomyocyte differentiation of human embryonic stem cells
Zara Melkoumian,Jennifer L. Weber,David M. Weber,Andrei Gennadyevich Fadeev,Yue Zhou,Paula Dolley-Sonneville,Jiwei Yang,Liqun Qiu,Catherine A. Priest,Christopher B. Shogbon,Arthur W. Martin,Jodelle Karen Nelson,Peter West,James P. Beltzer,Santona Pal,Ralph Brandenberger +15 more
TL;DR: Synthetic peptide-acrylate surfaces (PAS) are described that support self-renewal of hESCs in chemically defined, xeno-free medium and cell morphology and phenotypic marker expression were similar for cells cultured on PAS or Matrigel.
Journal ArticleDOI
Microwell-mediated control of embryoid body size regulates embryonic stem cell fate via differential expression of WNT5a and WNT11
Yu-Shik Hwang,Bong Geun Chung,Daniel Ortmann,Nobuaki Hattori,Hannes-Christian Moeller,Ali Khademhosseini +5 more
TL;DR: The data suggest that EB size could be an important parameter in ES cell fate specification via differential gene expression of members of the noncanonical WNT pathway, according to EB size.