New frontiers in human cell biology and medicine: Can pluripotent stem cells deliver?

Human pluripotent stem cells provide enormous opportunities to treat disease using cell therapy. But human stem cells can also drive biomedical and cell biological discoveries in a human model system, which can be directly linked to understanding disease or developing new therapies. Finally, rigorous scientific studies of these cells can and should inform the many science and medical policy issues that confront the translation of these technologies to medicine. In this paper, I discuss these issues using amyotrophic lateral sclerosis as an example.

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Telomere length set point regulation in human pluripotent stem cells critically depends on the shelterin protein TPP1

Molecular Biology of the Cell ◽

10.1091/mbc.e19-08-0447 ◽

2020 ◽

Vol 31 (23) ◽

pp. 2583-2596

Author(s):

John M. Boyle ◽

Kelsey M. Hennick ◽

Samuel G. Regalado ◽

Jacob M. Vogan ◽

Xiaozhu Zhang ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Telomere Length ◽

Pluripotent Stem Cells ◽

Human Stem Cells ◽

Set Point ◽

Hypomorphic Allele ◽

Point Control ◽

Set Point Control ◽

Stem Cell Lines

To better understand telomere length set point control in human stem cells, we generated knockout stem cell lines for TPP1 and contrasted their phenotypes with those of homozygous TPP1 L104A mutant stem cells. This comparison reveals that TPP1 L104A is not a hypomorphic allele but formally establishes TPP1 L104 as a dissociation of function mutant.

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Systematic gene tagging using CRISPR/Cas9 in human stem cells to illuminate cell organization

10.1101/123042 ◽

2017 ◽

Cited By ~ 1

Author(s):

Brock Roberts ◽

Amanda Haupt ◽

Andrew Tucker ◽

Tanya Grancharova ◽

Joy Arakaki ◽

...

Keyword(s):

Stem Cells ◽

Quality Control ◽

Genome Editing ◽

Cell Biology ◽

Cellular Structures ◽

Human Stem Cells ◽

Alpha Tubulin ◽

Junction Protein ◽

Endogenous Proteins ◽

Clonal Lines

AbstractWe present a CRISPR/Cas9 genome editing strategy to systematically tag endogenous proteins with fluorescent tags in human inducible pluripotent stem cells. To date we have generated multiple human iPSC lines with GFP tags for 10 proteins representing key cellular structures. The tagged proteins include alpha tubulin, beta actin, desmoplakin, fibrillarin, lamin B1, non-muscle myosin heavy chain IIB, paxillin, Sec61 beta, tight junction protein ZO1, and Tom20. Our genome editing methodology using Cas9 ribonuclear protein electroporation and fluorescence-based enrichment of edited cells resulted in <0.1-24% HDR across all experiments. Clones were generated from each edited population and screened for precise editing. ∼25% of the clones contained precise mono-allelic edits at the targeted locus. Furthermore, 92% (36/39) of expanded clonal lines satisfied key quality control criteria including genomic stability, appropriate expression and localization of the tagged protein, and pluripotency. Final clonal lines corresponding to each of the 10 cellular structures are now available to the research community. The data described here, including our editing protocol, genetic screening, quality control assays, and imaging observations, can serve as an initial resource for genome editing in cell biology and stem cell research.

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3. Personalized pluripotent stem cells

10.1093/actrade/9780198869290.003.0003 ◽

2021 ◽

pp. 39-51

Author(s):

Jonathan Slack

Keyword(s):

Stem Cells ◽

Pluripotent Stem Cells ◽

Cell Biology ◽

Nuclear Transplantation ◽

Stem Cell Biology ◽

Normal Cells ◽

Specific Patient ◽

No Formation ◽

Induced Pluripotent ◽

The Individual

‘Personalized pluripotent stem cells’ discusses cloning and its connection to stem cell biology. Somatic cell nuclear transplantation into oocytes can make personalized pluripotent stem cells as a perfect genetic match to a specific patient that provoke no immune rejection on grafting. Because this procedure involves generation of cells but no formation of an actual cloned individual, it has become known as human therapeutic cloning. Induced pluripotent stem cells (iPS cells) are made by introducing a few specific genes into normal cells. They are also a perfect genetic match to the individual donating the normal cells and because they are easy to make are now the preferred source.

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Advances in Pluripotent Stem Cells: History, Mechanisms, Technologies, and Applications

Stem Cell Reviews and Reports ◽

10.1007/s12015-019-09935-x ◽

2019 ◽

Vol 16 (1) ◽

pp. 3-32 ◽

Cited By ~ 22

Author(s):

Gele Liu ◽

Brian T. David ◽

Matthew Trawczynski ◽

Richard G. Fessler

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Regenerative Medicine ◽

Pluripotent Stem Cells ◽

Cell Biology ◽

Ethical Issues ◽

Three Dimensional ◽

Future Research ◽

Cell Technology

AbstractOver the past 20 years, and particularly in the last decade, significant developmental milestones have driven basic, translational, and clinical advances in the field of stem cell and regenerative medicine. In this article, we provide a systemic overview of the major recent discoveries in this exciting and rapidly developing field. We begin by discussing experimental advances in the generation and differentiation of pluripotent stem cells (PSCs), next moving to the maintenance of stem cells in different culture types, and finishing with a discussion of three-dimensional (3D) cell technology and future stem cell applications. Specifically, we highlight the following crucial domains: 1) sources of pluripotent cells; 2) next-generation in vivo direct reprogramming technology; 3) cell types derived from PSCs and the influence of genetic memory; 4) induction of pluripotency with genomic modifications; 5) construction of vectors with reprogramming factor combinations; 6) enhancing pluripotency with small molecules and genetic signaling pathways; 7) induction of cell reprogramming by RNA signaling; 8) induction and enhancement of pluripotency with chemicals; 9) maintenance of pluripotency and genomic stability in induced pluripotent stem cells (iPSCs); 10) feeder-free and xenon-free culture environments; 11) biomaterial applications in stem cell biology; 12) three-dimensional (3D) cell technology; 13) 3D bioprinting; 14) downstream stem cell applications; and 15) current ethical issues in stem cell and regenerative medicine. This review, encompassing the fundamental concepts of regenerative medicine, is intended to provide a comprehensive portrait of important progress in stem cell research and development. Innovative technologies and real-world applications are emphasized for readers interested in the exciting, promising, and challenging field of stem cells and those seeking guidance in planning future research direction.

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ALS-related FUS mutations alter axon growth in motoneurons and affect HuD/ELAVL4 and FMRP activity

Communications Biology ◽

10.1038/s42003-021-02538-8 ◽

2021 ◽

Vol 4 (1) ◽

Author(s):

Maria Giovanna Garone ◽

Nicol Birsa ◽

Maria Rosito ◽

Federico Salaris ◽

Michela Mochi ◽

...

Keyword(s):

Stem Cells ◽

Pluripotent Stem Cells ◽

Rna Binding ◽

Rna Binding Protein ◽

Axon Growth ◽

Motoneuron Disease ◽

Transcript Levels ◽

Protein Levels ◽

Induced Pluripotent ◽

Lateral Sclerosis

AbstractMutations in the RNA-binding protein (RBP) FUS have been genetically associated with the motoneuron disease amyotrophic lateral sclerosis (ALS). Using both human induced pluripotent stem cells and mouse models, we found that FUS-ALS causative mutations affect the activity of two relevant RBPs with important roles in neuronal RNA metabolism: HuD/ELAVL4 and FMRP. Mechanistically, mutant FUS leads to upregulation of HuD protein levels through competition with FMRP for HuD mRNA 3’UTR binding. In turn, increased HuD levels overly stabilize the transcript levels of its targets, NRN1 and GAP43. As a consequence, mutant FUS motoneurons show increased axon branching and growth upon injury, which could be rescued by dampening NRN1 levels. Since similar phenotypes have been previously described in SOD1 and TDP-43 mutant models, increased axonal growth and branching might represent broad early events in the pathogenesis of ALS.

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Biotechnology – opportunities for Europe

BSAP Occasional Publication ◽

10.1017/s0263967x00040337 ◽

2004 ◽

Vol 31 ◽

pp. 135-139

Author(s):

C. Warkup

Keyword(s):

Stem Cells ◽

Food Production ◽

New Technologies ◽

Human Medicine ◽

Early Years ◽

The Other ◽

Human Stem Cells ◽

The World ◽

Impartial Observer ◽

The Many

The title of this paper, as proposed by the meeting organisers, implies that Europe is different when it comes to biotechnology. In the early years of the 21st Century, even an impartial observer would agree that Europe differs from most of the rest of the world in its attitudes to at least one biotechnology – Genetically Modified (GM) crops. On the other hand, parts of Europe are seen as relatively enthusiastic about applications of biotechnology in human medicine. Take for instance, the UK's stance on research with human stem cells. Do these differences reflect permanent differences or merely a more cautious approach in Europe to the adoption of biotechnology in food production? Does this matter to pig producers?This paper seeks to give a broad and shallow overview of the opportunities for developments in biotechnology to impact on pig production. It will consider which of the many potential new technologies, if they were available now, might be acceptable in Europe and what might be the consequences of failure to access technologies that others exploit.

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