Culture System of Bile Duct-Like Cystic Structures Derived from Human-Inducible Pluripotent Stem Cells

2018 ◽  
pp. 143-153
Author(s):  
Akihide Kamiya ◽  
Kazuya Anzai ◽  
Kota Tsuruya ◽  
Hiromi Chikada
Keyword(s):  
Stem Cells ◽  
Bile Duct ◽  
Pluripotent Stem Cells ◽  
Culture System

scholarly journals Chemically defined and xeno-free culture condition for human extended pluripotent stem cells

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Bei Liu ◽  
Shi Chen ◽  
Yaxing Xu ◽  
Yulin Lyu ◽  
Jinlin Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Culture Condition ◽  
Human Fibroblast ◽  
Culture System ◽  
Disease Modeling ◽  
Directed Differentiation

AbstractExtended pluripotent stem (EPS) cells have shown great applicative potentials in generating synthetic embryos, directed differentiation and disease modeling. However, the lack of a xeno-free culture condition has significantly limited their applications. Here, we report a chemically defined and xeno-free culture system for culturing and deriving human EPS cells in vitro. Xeno-free human EPS cells can be long-term and genetically stably maintained in vitro, as well as preserve their embryonic and extraembryonic developmental potentials. Furthermore, the xeno-free culturing system also permits efficient derivation of human EPS cells from human fibroblast through reprogramming. Our study could have broad utility in future applications of human EPS cells in biomedicine.

Chemically defined and growth-factor-free culture system for the expansion and derivation of human pluripotent stem cells

2018 ◽  
Vol 2 (3) ◽  
pp. 173-182 ◽  
Author(s):  
Shin-ya Yasuda ◽  
Tatsuhiko Ikeda ◽  
Hosein Shahsavarani ◽  
Noriko Yoshida ◽  
Bhavana Nayer ◽  
...  
Keyword(s):  
Stem Cells ◽  
Growth Factor ◽  
Pluripotent Stem Cells ◽  
Culture System ◽  
Human Pluripotent Stem Cells

Scaling up a chemically-defined aggregate-based suspension culture system for neural commitment of human pluripotent stem cells

2016 ◽  
Vol 11 (12) ◽  
pp. 1628-1638 ◽  
Author(s):  
Cláudia C. Miranda ◽  
Tiago G. Fernandes ◽  
M. Margarida Diogo ◽  
Joaquim M.S. Cabral
Keyword(s):  
Stem Cells ◽  
Suspension Culture ◽  
Pluripotent Stem Cells ◽  
Culture System ◽  
Scaling Up ◽  
Human Pluripotent Stem Cells

scholarly journals Ascorbic Acid Can Promote the Generation and Expansion of Neuroepithelial-Like Stem Cells Derived from HiPS/ES Cells Under Chemically Defined Conditions Through Promoting Collagen Synthesis

2020 ◽  
Author(s):  
Rui Bai ◽  
Yun Chang ◽  
Amina Saleem ◽  
Fujian Wu ◽  
Lei Tian ◽  
...  
Keyword(s):  
Stem Cells ◽  
Ascorbic Acid ◽  
Pluripotent Stem Cells ◽  
Culture System ◽  
Serum Free Medium ◽  
Cord Injury ◽  
Induction System ◽  
Defined Culture

Abstract Introduction: Spinal cord injury (SCI) is a neurological, medically incurable disorder. Human pluripotent stem cells (hPSCs) have the potential to generate neural stem/progenitor cells (NS/PCs) which hold promise in therapy for SCI by transplantation. In our study, we aimed to establish a chemically defined culture system by using serum-free medium and ascorbic acid (AA) to generation and expansion of long-term self-renewing neuroepithelial-like stem cells (lt-NES cells) differentiated from hPSCs effectively and stably. Methods: We induce hESC/iPSC to neurospheres by using a newly established induction system in vitro in our study. And lt-NES cells derived from hESCs/iPSCs-neurospheres using two induction systems, including conventional N2 medium with gelatin-coated (coated) and N2+AA medium without pre-coated (AA) were characterized by reverse transcription-polymerase chain reaction (RT-PCR) analysis and immunocytochemistry staining. Subsequently, lt-NES cells were induced to neurons and the microelectrode array (MEA) recording system was used to evaluate the functionality of neurons differentiated from lt-NES cells. Moreover, the mechanism of AA-induced lt-NES cells was explored through RNA-seq and the use of inhibitors. Results: HESCs/iPSCs were efficiently induced to neurospheres by using a newly established induction system in vitro. And lt-NES cells derived from hESCs/iPSCs-neurospheres using two induction system (coated vs AA) both expressed neural pluripotency-associated genes PAX6, NESTIN, SOX1, SOX2. After long-term cultivation, we found that they both can maintain the long-term expansion for more than a dozen generations while maintaining neuropluripotency. Moreover, the lt-NES cells retain the ability to differentiate into general functional neurons that highly express β-tubulin. We also demonstrated that AA promotes the generation and long-term expansion of lt-NES cells by promoting collagen synthesis via the MEK-ERK1/2 pathways. Conclusions: Taken together, this new chemically defined culture system is stable and effective to generate and culture the lt-NES cells induced by hESCs/iPSCs using serum-free medium combined with ascorbic acid (AA). The lt-NES cells under this culture system can maintain the long-term expansion and neural pluripotency, with the potential to differentiate into functional neurons. Keywords: Spinal cord injury, Neurospheres, Ascorbic acid, lt-NES cells, Human pluripotent stem cells.

Generation of human hair-bearing skin organoids from stem cells

2020 ◽  
Author(s):  
Jiyoon Lee ◽  
Karl Koehler
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Human Hair ◽  
Culture System ◽  
Disease Modeling ◽  
Three Dimensional ◽  
Skin Tissue ◽  
Organoid Culture ◽  
Skin Biology

Abstract Skin is a complex and vulnerable tissue that it is challenging to reconstitute once damaged. Here, we describe a three-dimensional organoid culture system that can generate fully stratified skin with its appendages from human pluripotent stem cells. This in vitro-based skin organoid culture system will benefit investigations into basic skin biology and disease modeling, as well as translational efforts to reconstruct or regenerate skin tissue.

scholarly journals Generation of cortical neurons through large-scale expanding neuroepithelial stem cell from human pluripotent stem cells

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Shumei Zhao ◽  
Kui Duan ◽  
Zongyong Ai ◽  
Baohua Niu ◽  
Yanying Chen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Suspension Culture ◽  
Pluripotent Stem Cells ◽  
Cortical Neurons ◽  
Culture System ◽  
Large Scale ◽  
Disease Modeling ◽  
High Quality ◽  
Differentiation Potential

Abstract Background Considerable progress has been made in converting human pluripotent stem cells (hPSCs) into cortical neurons for disease modeling and regenerative medicine. However, these procedures are hard to provide sufficient cells for their applications. Using a combination of small-molecules and growth factors, we previously identified one condition which can rapidly induce hPSCs into neuroepithelial stem cells (NESCs). Here, we developed a scalable suspension culture system, which largely yields high-quality NESC-spheres and subsequent cortical neurons. Methods The NESC medium was first optimized, and the suspension culture system was then enlarged from plates to stirred bioreactors for large-scale production of NESC-spheres by a stirring speed of 60 rpm. During the expansion, the quality of NESC-spheres was evaluated. The differentiation potential of NESC-spheres into cortical neurons was demonstrated by removing bFGF and two pathway inhibitors from the NESC medium. Cellular immunofluorescence staining, global transcriptome, and single-cell RNA sequencing analysis were used to identify the characteristics, identities, purities, or homogeneities of NESC-spheres or their differentiated cells, respectively. Results The optimized culture system is more conducive to large-scale suspension production of NESCs. These largely expanded NESC-spheres maintain unlimited self-renewal ability and NESC state by retaining their uniform sizes, high cell vitalities, and robust expansion abilities. After long-term expansion, NESC-spheres preserve high purity, homogeneity, and normal diploid karyotype. These expanded NESC-spheres on a large scale have strong differentiation potential and effectively produce mature cortical neurons. Conclusions We developed a serum-free, defined, and low-cost culture system for large-scale expansion of NESCs in stirred suspension bioreactors. The stable and controllable 3D system supports long-term expansion of high-quality and homogeneous NESC-spheres. These NESC-spheres can be used to efficiently give rise to cortical neurons for cell therapy, disease modeling, and drug screening in future.

scholarly journals Establishment of automated culture system for murine induced pluripotent stem cells

2012 ◽  
Vol 12 (1) ◽  
pp. 81 ◽  
Author(s):  
Hiroyuki Koike ◽  
Koji Kubota ◽  
Keisuke Sekine ◽  
Takanori Takebe ◽  
Rie Ouchi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Culture System ◽  
Induced Pluripotent
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