Complex Organ Construction from Human Pluripotent Stem Cells for Biological Research and Disease Modeling with New Emerging Techniques

Human pluripotent stem cells (hPSCs) are grouped into two cell types; embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs). hESCs have provided multiple powerful platforms to study human biology, including human development and diseases; however, there were difficulties in the establishment of hESCs from human embryo and concerns over its ethical issues. The discovery of hiPSCs has expanded to various applications in no time because hiPSCs had already overcome these problems. Many hPSC-based studies have been performed using two-dimensional monocellular culture methods at the cellular level. However, in many physiological and pathophysiological conditions, intra- and inter-organ interactions play an essential role, which has hampered the establishment of an appropriate study model. Therefore, the application of recently developed technologies, such as three-dimensional organoids, bioengineering, and organ-on-a-chip technology, has great potential for constructing multicellular tissues, generating the functional organs from hPSCs, and recapitulating complex tissue functions for better biological research and disease modeling. Moreover, emerging techniques, such as single-cell transcriptomics, spatial transcriptomics, and artificial intelligence (AI) allowed for a denser and more precise analysis of such heterogeneous and complex tissues. Here, we review the applications of hPSCs to construct complex organs and discuss further prospects of disease modeling and drug discovery based on these PSC-derived organs.

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Pancreatic Progenitors and Organoids as a Prerequisite to Model Pancreatic Diseases and Cancer

Stem Cells International ◽

10.1155/2019/9301382 ◽

2019 ◽

Vol 2019 ◽

pp. 1-11 ◽

Cited By ~ 3

Author(s):

Meike Hohwieler ◽

Martin Müller ◽

Pierre-Olivier Frappart ◽

Sandra Heller

Keyword(s):

Stem Cells ◽

Pluripotent Stem Cells ◽

Disease Modeling ◽

Embryonic Stem ◽

Cell Types ◽

Genetically Engineered ◽

Hereditary Diseases ◽

Pancreatic Diseases ◽

Pancreatic Progenitors

Embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) are characterized by their unique capacity to stepwise differentiate towards any particular cell type in an adult organism. Pluripotent stem cells provide a beneficial platform to model hereditary diseases and even cancer development. While the incidence of pancreatic diseases such as diabetes and pancreatitis is increasing, the understanding of the underlying pathogenesis of particular diseases remains limited. Only a few recent publications have contributed to the characterization of human pancreatic development in the fetal stage. Hence, most knowledge of pancreatic specification is based on murine embryology. Optimizing and understanding current in vitro protocols for pancreatic differentiation of ESCs and iPSCs constitutes a prerequisite to generate functional pancreatic cells for better disease modeling and drug discovery. Moreover, human pancreatic organoids derived from pluripotent stem cells, organ-restricted stem cells, and tumor samples provide a powerful technology to model carcinogenesis and hereditary diseases independent of genetically engineered mouse models. Herein, we summarize recent advances in directed differentiation of pancreatic organoids comprising endocrine cell types. Beyond that, we illustrate up-and-coming applications for organoid-based platforms.

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Advances in Stem Cell Technologies for Disease Therapy: A Current Review

Annals of Advanced Biomedical Sciences ◽

10.23880/aabsc-16000107 ◽

2018 ◽

Vol 1 (1) ◽

Author(s):

Nadiya Patel

Keyword(s):

Stem Cells ◽

Pluripotent Stem Cells ◽

Ethical Issues ◽

Therapeutic Potential ◽

Embryonic Stem ◽

Cell Types ◽

Current Review ◽

Disease Therapy ◽

Induced Pluripotent ◽

A Current

Stem cells have the capability of differentiating into limitless cell types, alongside the function of exceptional proliferative capacity. There are three main types of stem cells: embryonic stem cells (ESCs), induced pluripotent stem cells (IPSCs) and mesenchymal stem cells (MSCs). ESCs are highly versatile and hold great therapeutic potential but have great ethical barriers and considerations that are yet to be overcome. IPSCs have become increasingly popular within research as they are not restrained by any ethical issues and do not require approval for their usage. The aim of this review was to expand on the background and therapeutic potential of ESCs and IPSCs whilst linking this to their use within disease therapy with a specific focus on ethics, tumorigenesis and survivability. The analysis found some conflicting results and a delay in the advance of overcoming the problems of tumorigenesis and survivability of stem cells. Both stem cells types have shown good efficacy but do also come with their disadvantages.

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Prospect of Human Pluripotent Stem Cell-Derived Neural Crest Stem Cells in Clinical Application

Stem Cells International ◽

10.1155/2016/7695836 ◽

2016 ◽

Vol 2016 ◽

pp. 1-11 ◽

Cited By ~ 7

Author(s):

Qian Zhu ◽

Qiqi Lu ◽

Rong Gao ◽

Tong Cao

Keyword(s):

Stem Cells ◽

Nervous System ◽

Neural Crest ◽

Clinical Application ◽

Pluripotent Stem Cells ◽

Embryonic Stem ◽

Hirschsprung Disease ◽

Cell Types ◽

Human Pluripotent Stem Cells ◽

Neural Crest Stem Cells

Neural crest stem cells (NCSCs) represent a transient and multipotent cell population that contributes to numerous anatomical structures such as peripheral nervous system, teeth, and cornea. NCSC maldevelopment is related to various human diseases including pigmentation abnormalities, disorders affecting autonomic nervous system, and malformations of teeth, eyes, and hearts. As human pluripotent stem cells including human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) can serve as an unlimited cell source to generate NCSCs, hESC/hiPSC-derived NCSCs can be a valuable tool to study the underlying mechanisms of NCSC-associated diseases, which paves the way for future therapies for these abnormalities. In addition, hESC/hiPSC-derived NCSCs with the capability of differentiating to various cell types are highly promising for clinical organ repair and regeneration. In this review, we first discuss NCSC generation methods from human pluripotent stem cells and differentiation mechanism of NCSCs. Then we focus on the clinical application potential of hESC/hiPSC-derived NCSCs on peripheral nerve injuries, corneal blindness, tooth regeneration, pathological melanogenesis, Hirschsprung disease, and cardiac repair and regeneration.

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Identification of an epigenetic signature in human induced pluripotent stem cells using a linear machine learning model

Human Cell ◽

10.1007/s13577-020-00446-3 ◽

2020 ◽

Vol 34 (1) ◽

pp. 99-110

Author(s):

Koichiro Nishino ◽

Ken Takasawa ◽

Kohji Okamura ◽

Yoshikazu Arai ◽

Asato Sekiya ◽

...

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Ethical Issues ◽

Embryonic Stem ◽

Learning Model ◽

Carcinoma Cells ◽

Biological Research ◽

Machine Learning Model ◽

Induced Pluripotent

AbstractThe use of human induced pluripotent stem cells (iPSCs), used as an alternative to human embryonic stem cells (ESCs), is a potential solution to challenges, such as immune rejection, and does not involve the ethical issues concerning the use of ESCs in regenerative medicine, thereby enabling developments in biological research. However, comparative analyses from previous studies have not indicated any specific feature that distinguishes iPSCs from ESCs. Therefore, in this study, we established a linear classification-based learning model to distinguish among ESCs, iPSCs, embryonal carcinoma cells (ECCs), and somatic cells on the basis of their DNA methylation profiles. The highest accuracy achieved by the learned models in identifying the cell type was 94.23%. In addition, the epigenetic signature of iPSCs, which is distinct from that of ESCs, was identified by component analysis of the learned models. The iPSC-specific regions with methylation fluctuations were abundant on chromosomes 7, 8, 12, and 22. The method developed in this study can be utilized with comprehensive data and widely applied to many aspects of molecular biology research.

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Comprehensive analysis of prime editing outcomes in human embryonic stem cells

10.1101/2021.04.12.439533 ◽

2021 ◽

Author(s):

Omer Habib ◽

Gizem Habib ◽

Gue-ho Hwang ◽

Sangsu Bae

Keyword(s):

Stem Cells ◽

Genome Editing ◽

Pluripotent Stem Cells ◽

Disease Modeling ◽

Cytidine Deaminase ◽

Embryonic Stem ◽

Cell Types ◽

Great Promise ◽

Nucleotide Substitutions ◽

Base Editing

Prime editing is a versatile and precise genome editing technique that can directly copy desired genetic modifications into target DNA sites without the need for donor DNA. This technique holds great promise for the analysis of gene function, disease modeling, and the correction of pathogenic mutations in clinically relevant cells such as human pluripotent stem cells (hPSCs). Here we comprehensively tested prime editing in hPSCs by generating a doxycycline-inducible prime editing platform. Prime editing successfully induced all types of nucleotide substitutions and small insertions and deletions, similar to observations in other human cell types. Moreover, we compared prime editing and base editing for correcting a disease-related mutation in induced pluripotent stem cells derived form a patient with α 1-antitrypsin (A1AT) deficiency. Finally, whole-genome sequencing showed that, unlike the cytidine deaminase domain of cytosine base editors, the reverse transcriptase domain of a prime editor does not lead to guide RNA-independent off-target mutations in the genome. Our results demonstrate that prime editing in hPSCs has great potential for complementing previously developed CRISPR genome editing tools.

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Questionnaire for evaluating information, knowledge, and attitudes on donation, storage, and application of induced pluripotent stem cells

Genetika ◽

10.2298/gensr2102813r ◽

2021 ◽

Vol 53 (2) ◽

pp. 813-823

Author(s):

Sanja Rascanin ◽

Mirjana Jovanovic ◽

Dejan Stevanovic ◽

Nemanja Rancic

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Drug Testing ◽

Disease Modeling ◽

Ethical Issues ◽

Embryonic Stem ◽

Knowledge And Attitudes ◽

Induced Pluripotent

The discovery of Induced Pluripotent Stem Cells (iPSCs) opened the possibilities for reprogramming adult somatic cells back to a pluripotent state in vitro by inducing a forced expression of specific transcription factors. Thus, iPSCs might have potential application in regenerative medicine, transplantation, avoidance of tissue rejection, disease modeling, and drug testing. Because of apparent ethical issues connected with donation and derivation of biomaterials, iPSCs are considered as a research alternative to ethically highly disputed Embryonic Stem Cells (ESCs). Objective: The aim of this paper was to describe the development of a questionnaire for evaluating information, knowledge, and attitudes on donation, storage, and application of iPSCs (i.e., the QIPSC). We performed a prospective qualitative study based on the development, validation and reliability testing of the QIPSC. The study included 122 respondents and the final version of the QIPSC with 34 items. The reliability analysis for part of information and knowledge of respondents according to iPSCs was then performed with the questions included in this two-component model and obtained a Cronbach's alpha value of 0.783 and 0.870, respectively. It has been shown that the range of correct answers to questions in part of knowledge of respondents according to iPSCs was from 17.2-63.1%. The results of our study show that the QIPSC was a unique, reliable, and valid questionnaire for assessing the level of information, knowledge, and attitudes on donation, storage, and application of iPSCs.

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Induced Pluripotent Stem Cells: Generation Strategy and Epigenetic Mystery behind Reprogramming

Stem Cells International ◽

10.1155/2016/8415010 ◽

2016 ◽

Vol 2016 ◽

pp. 1-11 ◽

Cited By ~ 5

Author(s):

Pengfei Ji ◽

Sasicha Manupipatpong ◽

Nina Xie ◽

Yujing Li

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Clinical Medicine ◽

Ethical Issues ◽

Somatic Cells ◽

Embryonic Stem ◽

Cell Types ◽

Self Renewal ◽

Induced Pluripotent

Possessing the ability of self-renewal with immortalization and potential for differentiation into different cell types, stem cells, particularly embryonic stem cells (ESC), have attracted significant attention since their discovery. As ESC research has played an essential role in developing our understanding of the mechanisms underlying reproduction, development, and cell (de)differentiation, significant efforts have been made in the biomedical study of ESC in recent decades. However, such studies of ESC have been hampered by the ethical issues and technological challenges surrounding them, therefore dramatically inhibiting the potential applications of ESC in basic biomedical studies and clinical medicine. Induced pluripotent stem cells (iPSCs), generated from the reprogrammed somatic cells, share similar characteristics including but not limited to the morphology and growth of ESC, self-renewal, and potential differentiation into various cell types. The discovery of the iPSC, unhindered by the aforementioned limitations of ESC, introduces a viable alternative to ESC. More importantly, the applications of iPSC in the development of disease models such as neurodegenerative disorders greatly enhance our understanding of the pathogenesis of such diseases and also facilitate the development of clinical therapeutic strategies using iPSC generated from patient somatic cells to avoid an immune rejection. In this review, we highlight the advances in iPSCs generation methods as well as the mechanisms behind their reprogramming. We also discuss future perspectives for the development of iPSC generation methods with higher efficiency and safety.

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Interspecies Chimeric Conditions Affect the Developmental Rate of Human Pluripotent Stem Cells

10.1101/2020.09.12.293357 ◽

2020 ◽

Author(s):

Jared Brown ◽

Christopher Barry ◽

Matthew T. Schmitz ◽

Cara Argus ◽

Jennifer M. Bolin ◽

...

Keyword(s):

Stem Cells ◽

Pluripotent Stem Cells ◽

Embryonic Stem ◽

Cell Types ◽

In Utero ◽

Human Pluripotent Stem Cells ◽

Human Organs ◽

Species Specific ◽

Dose Dependent

ABSTRACTHuman pluripotent stem cells hold significant promise for regenerative medicine. However, long differentiation protocols and immature characteristics of stem cell-derived cell types remain challenges to the development of many therapeutic applications. In contrast to the slow differentiation of human stem cells in vitro that mirrors a nine-month gestation period, mouse stem cells develop according to a much faster three-week gestation timeline. Here, we tested if co-differentiation with mouse pluripotent stem cells could accelerate the differentiation speed of human embryonic stem cells. Following a six-week RNA-sequencing time course of neural differentiation, we identified 929 human genes that were upregulated earlier and 535 genes that exhibited earlier peaked expression profiles in chimeric cell cultures than in human cell cultures alone. Genes with accelerated upregulation were significantly enriched in Gene Ontology terms associated with neurogenesis, neuron differentiation and maturation, and synapse signaling. Moreover, chimeric mixed samples correlated with in utero human embryonic samples earlier than human cells alone, and acceleration was dose-dependent on human-mouse co-culture ratios. Differences in the timing and expression levels of genes corresponding to neuron cell types and brain region identity under chimeric conditions were also observed. The altered developmental rates and lineage outcomes described in this report have implications for accelerating human stem cell differentiation and the use of interspecies chimeric embryos in developing human organs for transplantation.Author SummaryHuman pluripotent stem cells often require long in vitro protocols to form mature cell types of clinical relevance for potential regenerative therapies, a ramification of a nine-month developmental clock in utero that also runs ex utero. What controls species-specific developmental time and whether the timer is amenable to acceleration is unknown. Further, interspecies chimeric embryos are increasingly being created to study early human development or explore the potential growth of human organs for transplantation. How the conflicting developmental speeds of cells from different species co-differentiating together affect each other is not understood. Here, using genome-wide transcriptional analysis of RNA-sequencing time courses, we show that 1) co-differentiating human embryonic stem cells intermixed with mouse stem cells accelerated elements of human developmental programs, 2) the acceleration was dose-dependent on the proportion of mouse cells, and 3) human cells in chimeric samples correlated to in utero samples earlier than human only samples. Our results provide evidence that some components of species-specific developmental clocks may be susceptible to acceleration.

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Mini Review; Differentiation of Human Pluripotent Stem Cells into Oocytes

Current Stem Cell Research & Therapy ◽

10.2174/1574888x15666200116100121 ◽

2020 ◽

Vol 15 (4) ◽

pp. 301-307 ◽

Cited By ~ 3

Author(s):

Gaifang Wang ◽

Maryam Farzaneh

Keyword(s):

Stem Cells ◽

Pluripotent Stem Cells ◽

Embryonic Stem ◽

Human Pluripotent Stem Cells ◽

Human Oocyte ◽

Differentiation Potential ◽

Cell Lineages ◽

Cell Based Therapy ◽

Induced Pluripotent

Primary Ovarian Insufficiency (POI) is one of the main diseases causing female infertility that occurs in about 1% of women between 30-40 years of age. There are few effective methods for the treatment of women with POI. In the past few years, stem cell-based therapy as one of the most highly investigated new therapies has emerged as a promising strategy for the treatment of POI. Human pluripotent stem cells (hPSCs) can self-renew indefinitely and differentiate into any type of cell. Human Embryonic Stem Cells (hESCs) as a type of pluripotent stem cells are the most powerful candidate for the treatment of POI. Human-induced Pluripotent Stem Cells (hiPSCs) are derived from adult somatic cells by the treatment with exogenous defined factors to create an embryonic-like pluripotent state. Both hiPSCs and hESCs can proliferate and give rise to ectodermal, mesodermal, endodermal, and germ cell lineages. After ovarian stimulation, the number of available oocytes is limited and the yield of total oocytes with high quality is low. Therefore, a robust and reproducible in-vitro culture system that supports the differentiation of human oocytes from PSCs is necessary. Very few studies have focused on the derivation of oocyte-like cells from hiPSCs and the details of hPSCs differentiation into oocytes have not been fully investigated. Therefore, in this review, we focus on the differentiation potential of hPSCs into human oocyte-like cells.

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Characterization of Endoplasmic Reticulum (ER) in Human Pluripotent Stem Cells Revealed Increased Susceptibility to Cell Death upon ER Stress

Cells ◽

10.3390/cells9051078 ◽

2020 ◽

Vol 9 (5) ◽

pp. 1078

Author(s):

Tae Won Ha ◽

Ji Hun Jeong ◽

HyeonSeok Shin ◽

Hyun Kyu Kim ◽

Jeong Suk Im ◽

...

Keyword(s):

Stem Cells ◽

Endoplasmic Reticulum ◽

Er Stress ◽

Pluripotent Stem Cells ◽

Meta Analysis ◽

Embryonic Stem ◽

Structural Features ◽

Human Pluripotent Stem Cells ◽

Differentiated Cells ◽

Induced Apoptosis

Human pluripotent stem cells (hPSCs), such as embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), have a well-orchestrated program for differentiation and self-renewal. However, the structural features of unique proteostatic-maintaining mechanisms in hPSCs and their features, distinct from those of differentiated cells, in response to cellular stress remain unclear. We evaluated and compared the morphological features and stress response of hPSCs and fibroblasts. Compared to fibroblasts, electron microscopy showed simpler/fewer structures with fewer networks in the endoplasmic reticulum (ER) of hPSCs, as well as lower expression of ER-related genes according to meta-analysis. As hPSCs contain low levels of binding immunoglobulin protein (BiP), an ER chaperone, thapsigargin treatment sharply increased the gene expression of the unfolded protein response. Thus, hPSCs with decreased chaperone function reacted sensitively to ER stress and entered apoptosis faster than fibroblasts. Such ER stress-induced apoptotic processes were abolished by tauroursodeoxycholic acid, an ER-stress reliever. Hence, our results revealed that as PSCs have an underdeveloped structure and express fewer BiP chaperone proteins than somatic cells, they are more susceptible to ER stress-induced apoptosis in response to stress.

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