scholarly journals Screening Genes Promoting Exit from Naive Pluripotency Based on Genome-Scale CRISPR-Cas9 Knockout

2020 ◽  
Vol 2020 ◽  
pp. 1-15 ◽  
Author(s):  
Bin Yang ◽  
Junqi Kuang ◽  
Chuman Wu ◽  
Wenyi Zhou ◽  
Shuoji Zhu ◽  
...  
Keyword(s):  
Stem Cell ◽  
Regenerative Medicine ◽  
Large Scale ◽  
Gene Knockout ◽  
Embryonic Stem ◽  
Specific Cell ◽  
Medicine Research ◽  
Model Based ◽  
Key Genes ◽  
Genome Scale

Two of the main problems of stem cell and regenerative medicine are the exit of pluripotency and differentiation to functional cells or tissues. The answer to these two problems holds great value in the clinical translation of stem cell as well as regenerative medicine research. Although piling researches have revealed the truth about pluripotency maintenance, the mechanisms underlying pluripotent cell self-renewal, proliferation, and differentiation into specific cell lineages or tissues are yet to be defined. To this end, we took full advantage of a novel technology, namely, the genome-scale CRISPR-Cas9 knockout (GeCKO). As an effective way of introducing targeted loss-of-function mutations at specific sites in the genome, GeCKO is able to screen in an unbiased manner for key genes that promote exit from pluripotency in mouse embryonic stem cells (mESCs) for the first time. In this study, we successfully established a model based on GeCKO to screen the key genes in pluripotency withdrawal. Our strategies included lentiviral package and infection technology, lenti-Cas9 gene knockout technology, shRNA gene knockdown technology, next-generation sequencing, model-based analysis of genome-scale CRISPR-Cas9 knockout (MAGeCK analysis), GO analysis, and other methods. Our findings provide a novel approach for large-scale screening of genes involved in pluripotency exit and offer an entry point for cell fate regulation research.

RESEARCH FINDINGS

2011 ◽  
Vol 15 (12) ◽  
pp. 43-44
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Kawasaki Disease ◽  
Regenerative Medicine ◽  
Embryonic Stem Cell ◽  
Human Embryonic Stem Cell ◽  
Embryonic Stem ◽  
Medicine Research ◽  
Research Findings ◽  
Cell Study

Singapore Scientists Lead Human Embryonic Stem Cell Study to Advance Regenerative Medicine Research. Singapore Scientists Discover Genetic Link in Kawasaki Disease. Stem Cells Engineered to Kill Cancer.

scholarly journals Human Pluripotent Stem Cell Culture: Current Status, Challenges, and Advancement

2018 ◽  
Vol 2018 ◽  
pp. 1-17 ◽  
Author(s):  
Sushrut Dakhore ◽  
Bhavana Nayer ◽  
Kouichi Hasegawa
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Pluripotent Stem Cells ◽  
Large Scale ◽  
Embryonic Stem ◽  
Current Status ◽  
Specific Cell ◽  
Therapeutic Applications ◽  
Proliferative Ability

Over the past two decades, human embryonic stem cells (hESCs) have gained attention due to their pluripotent and proliferative ability which enables production of almost all cell types in the human body in vitro and makes them an excellent tool to study human embryogenesis and disease, as well as for drug discovery and cell transplantation therapies. Discovery of human-induced pluripotent stem cells (hiPSCs) further expanded therapeutic applications of human pluripotent stem cells (PSCs). hPSCs provide a stable and unlimited original cell source for producing suitable cells and tissues for downstream applications. Therefore, engineering the environment in which these cells are grown, for stable and quality-controlled hPSC maintenance and production, is one of the key factors governing the success of these applications. hPSCs are maintained in a particular niche using specific cell culture components. Ideally, the culture should be free of xenobiotic components to render hPSCs suitable for therapeutic applications. Substantial efforts have been put to identify effective components, and develop culture conditions and protocols, for their large-scale expansion without compromising on quality. In this review, we discuss different media, their components and functions, including specific requirements to maintain the pluripotent and proliferative ability of hPSCs. Understanding the role of culture components would enable the development of appropriate conditions to promote large-scale, quality-controlled expansion of hPSCs thereby increasing their potential applications.

scholarly journals The ACTCellerate initiative: large-scale combinatorial cloning of novel human embryonic stem cell derivatives

2008 ◽  
Vol 3 (3) ◽  
pp. 287-308 ◽  
Author(s):  
Michael D West ◽  
R Geoffrey Sargent ◽  
Jeff Long ◽  
Colleen Brown ◽  
Jing Song Chu ◽  
...  
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Human Embryonic Stem Cell ◽  
Large Scale ◽  
Embryonic Stem

Large-scale production of murine embryonic stem cell-derived osteoblasts and chondrocytes on microcarriers in serum-free media

Biomaterials ◽  
2011 ◽  
Vol 32 (26) ◽  
pp. 6006-6016 ◽  
Author(s):  
Roz Alfred ◽  
Jaymi T. Taiani ◽  
Roman J. Krawetz ◽  
Akihiro Yamashita ◽  
Derrick E. Rancourt ◽  
...  
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Large Scale ◽  
Embryonic Stem ◽  
Scale Production ◽  
Serum Free ◽  
Murine Embryonic Stem Cell ◽  
Large Scale Production ◽  
Free Media

scholarly journals Impedance-based cellular assays for regenerative medicine

2018 ◽  
Vol 373 (1750) ◽  
pp. 20170226 ◽  
Author(s):  
W. Gamal ◽  
H. Wu ◽  
I. Underwood ◽  
J. Jia ◽  
S. Smith ◽  
...  
Keyword(s):  
Stem Cell ◽  
Regenerative Medicine ◽  
Large Scale ◽  
Three Dimensional ◽  
Cell Renewal ◽  
Label Free ◽  
Impedance Tomography ◽  
Cellular Assays ◽  
Automation And Control

Therapies based on regenerative techniques have the potential to radically improve healthcare in the coming years. As a result, there is an emerging need for non-destructive and label-free technologies to assess the quality of engineered tissues and cell-based products prior to their use in the clinic. In parallel, the emerging regenerative medicine industry that aims to produce stem cells and their progeny on a large scale will benefit from moving away from existing destructive biochemical assays towards data-driven automation and control at the industrial scale. Impedance-based cellular assays (IBCA) have emerged as an alternative approach to study stem-cell properties and cumulative studies, reviewed here, have shown their potential to monitor stem-cell renewal, differentiation and maturation. They offer a novel method to non-destructively assess and quality-control stem-cell cultures. In addition, when combined with in vitro disease models they provide complementary insights as label-free phenotypic assays. IBCA provide quantitative and very sensitive results that can easily be automated and up-scaled in multi-well format. When facing the emerging challenge of real-time monitoring of three-dimensional cell culture dielectric spectroscopy and electrical impedance tomography represent viable alternatives to two-dimensional impedance sensing. This article is part of the theme issue ‘Designer human tissue: coming to a lab near you’.

scholarly journals The E3 Ligase Axotrophin/MARCH-7: Protein Expression Profiling of Human Tissues Reveals Links to Adult Stem Cells

2009 ◽  
Vol 58 (4) ◽  
pp. 301-308 ◽  
Author(s):  
Cristina A. Szigyarto ◽  
Paul Sibbons ◽  
Gill Williams ◽  
Mathias Uhlen ◽  
Su M. Metcalfe
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Protein Expression ◽  
Adult Stem Cells ◽  
Embryonic Stem ◽  
Cell Types ◽  
Null Mouse ◽  
Specific Cell ◽  
Direct Role ◽  
Neuronal Progenitor

Axotrophin/MARCH-7 was first identified in mouse embryonic stem cells as a neural stem cell gene. Using the axotrophin/MARCH-7 null mouse, we discovered profound effects on T lymphocyte responses, including 8-fold hyperproliferation and 5-fold excess release of the stem cell cytokine leukemia inhibitory factor (LIF). Our further discovery that axotrophin/MARCH-7 is required for targeted degradation of the LIF receptor subunit gp190 implies a direct role in the regulation of LIF signaling. Bioinformatics studies revealed a highly conserved RING-CH domain in common with the MARCH family of E3-ubiquitin ligases, and accordingly, axotrophin was renamed “MARCH-7.” To probe protein expression of human axotrophin/MARCH-7, we prepared antibodies against different domains of the protein. Each antibody bound its specific target epitope with high affinity, and immunohistochemistry cross-validated target specificity. Forty-eight human tissue types were screened. Epithelial cells stained strongly, with trophoblasts having the greatest staining. In certain tissues, specific cell types were selectively positive, including neurons and neuronal progenitor cells in the hippocampus and cerebellum, endothelial sinusoids of the spleen, megakaryocytes in the bone marrow, crypt stem cells of the small intestine, and alveolar macrophages in the lung. Approximately 20% of central nervous system neuropils were positive. Notably, axotrophin/MARCH-7 has an expression profile that is distinct from that of other MARCH family members. This manuscript contains online supplemental material at http://www.jhc.org . Please visit this article online to view these materials.

scholarly journals Stem Cells: Prospects and Potential Applications in Tanzania: A review

2020 ◽  
Vol 21 (2) ◽  
pp. 1-8
Author(s):  
Afadhali Denis Russa
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Regenerative Medicine ◽  
Large Scale ◽  
Adult Stem Cells ◽  
Treatment Options ◽  
Stem Cell Technology ◽  
African Countries ◽  
Cell Technology ◽  
Leading Role

Stem cell technology and its application in regenerative medicine is the future gateway for the treatment of most non-communicable diseases (NCDs). As the burden of NCDs continues to rises globally, regenerating the cells, tissues and organs will be the mainstream treatment option. The world is prepared for this intriguing but promising avenue of biomedical technology and medicine but Africa is grossly lagging far behind. African governments, universities, research and health institutions need to take a leading role in empowering and mainstreaming stem cell research.  Moreover, for Africa, there is a huge potential for translating stem cell technology into clinical treatments due to the fact that there are limited treatment options for life-threatening forms of NCDs.  Some African countries have well-developed stem cell facilities and large-scale stem cell therapy centers. The use of adult stem cells in liver failure, diabetes and cardiac infarcts has shown success in some African countries. The present work reviews the status, potential and future prospects of stem cell technology and regenerative medicine in Tanzania with particular emphasis on the adult stem cells applicability into the immediate use inpatient care.  The paper also reviews the available cell identification systems and markers and moral and ethical aspects of stem cell science necessary in the translational treatment regimens. 

scholarly journals Human therapeutic cloning, pitfalls and lack luster because of rapid developments in induced pluripotent stem cell technology

2014 ◽  
Vol 8 (1) ◽  
pp. 5-10
Author(s):  
Song Hua ◽  
Henry Chung ◽  
Kuldip Sidhu
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Regenerative Medicine ◽  
Somatic Cell ◽  
Pluripotent Stem Cells ◽  
Nuclear Transfer ◽  
Es Cells ◽  
Embryonic Stem ◽  
Therapeutic Cloning ◽  
Induced Pluripotent

AbstractBackground: Therapeutic cloning is the combination of somatic cell nuclear transfer (SCNT) and embryonic stem cell (ES) techniques to create specific ES cells that match those of a patient. Because ES cells derived by nuclear transfer (SCNT ES cells) are genetically identical to the donor, it will not generate rejection by the host’s immune system and thus therapeutically may be more acceptable. Induced pluripotent stem cells (iPS) are a type of pluripotent stem cell artificially derived from an adult somatic cell by inducing a forced expression of a set of specific pluripotent genes. In the past few years, rapid progress in reprogramming and iPS technology has been made, and it seems to shadow any progress made in SCNT programs.Objective: This review compares the application perspective of SCNT with that of iPS in regenerative medicine.Methods:We conducted a literature search using the MEDLINE (PubMed), Wiley InterScience, Springer, EBSCO, and Annual Reviews databases using the keywords “iPS”, “ES”, “SCNT” “induced pluripotent stem cells”, “embryonic stem cells”, “therapeutic cloning”, “regenerative medicine”, and “somatic cell nuclear transfer”. Only articles published in English were included in this review.Results: These two methods both have advantages and disadvantages. Nevertheless, by using SCNT to generate patient-specific cell lines, it eliminates complications by avoiding the use of viral vectors during iPS generation. Success in in vitro matured eggs from aged women and even differentiation of oocytes from germ stem cells will further enhance the application of SCNT in regenerative medicine.Conclusion: Human SCNT may be an appropriate mean of generating patient stem cell lines for clinical therapy in the near future.

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