Core-shell hydrogel microfiber-expanded pluripotent stem cell-derived lung progenitors applicable to lung reconstruction in vivo

Abstract Background Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM) are regarded as promising cell type for cardiac cell replacement therapy, but it is not known whether the developmental stage influences their persistence and functional integration in the host tissue, which are crucial for a long-term therapeutic benefit. To investigate this, we first tested the cell adhesion capability of murine iPSC-CM in vitro at three different time points during the differentiation process and then examined cell persistence and quality of electrical integration in the infarcted myocardium in vivo. Methods To test cell adhesion capabilities in vitro, iPSC-CM were seeded on fibronectin-coated cell culture dishes and decellularized ventricular extracellular matrix (ECM) scaffolds. After fixed periods of time, stably attached cells were quantified. For in vivo experiments, murine iPSC-CM expressing enhanced green fluorescent protein was injected into infarcted hearts of adult mice. After 6–7 days, viable ventricular tissue slices were prepared to enable action potential (AP) recordings in transplanted iPSC-CM and surrounding host cardiomyocytes. Afterwards, slices were lysed, and genomic DNA was prepared, which was then used for quantitative real-time PCR to evaluate grafted iPSC-CM count. Results The in vitro results indicated differences in cell adhesion capabilities between day 14, day 16, and day 18 iPSC-CM with day 14 iPSC-CM showing the largest number of attached cells on ECM scaffolds. After intramyocardial injection, day 14 iPSC-CM showed a significant higher cell count compared to day 16 iPSC-CM. AP measurements revealed no significant difference in the quality of electrical integration and only minor differences in AP properties between d14 and d16 iPSC-CM. Conclusion The results of the present study demonstrate that the developmental stage at the time of transplantation is crucial for the persistence of transplanted iPSC-CM. iPSC-CM at day 14 of differentiation showed the highest persistence after transplantation in vivo, which may be explained by a higher capability to adhere to the extracellular matrix.

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Modeling Type 1 Diabetes Using Pluripotent Stem Cell Technology

Frontiers in Endocrinology ◽

10.3389/fendo.2021.635662 ◽

2021 ◽

Vol 12 ◽

Author(s):

Kriti Joshi ◽

Fergus Cameron ◽

Swasti Tiwari ◽

Stuart I. Mannering ◽

Andrew G. Elefanty ◽

...

Keyword(s):

Type 1 Diabetes ◽

Stem Cell ◽

Genetic Variants ◽

Genetic Background ◽

Pluripotent Stem Cell ◽

Cell Types ◽

Polygenic Inheritance

Induced pluripotent stem cell (iPSC) technology is increasingly being used to create in vitro models of monogenic human disorders. This is possible because, by and large, the phenotypic consequences of such genetic variants are often confined to a specific and known cell type, and the genetic variants themselves can be clearly identified and controlled for using a standardized genetic background. In contrast, complex conditions such as autoimmune Type 1 diabetes (T1D) have a polygenic inheritance and are subject to diverse environmental influences. Moreover, the potential cell types thought to contribute to disease progression are many and varied. Furthermore, as HLA matching is critical for cell-cell interactions in disease pathogenesis, any model that seeks to test the involvement of particular cell types must take this restriction into account. As such, creation of an in vitro model of T1D will require a system that is cognizant of genetic background and enables the interaction of cells representing multiple lineages to be examined in the context of the relevant environmental disease triggers. In addition, as many of the lineages critical to the development of T1D cannot be easily generated from iPSCs, such models will likely require combinations of cell types derived from in vitro and in vivo sources. In this review we imagine what an ideal in vitro model of T1D might look like and discuss how the required elements could be feasibly assembled using existing technologies. We also examine recent advances towards this goal and discuss potential uses of this technology in contributing to our understanding of the mechanisms underlying this autoimmune condition.

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Human pluripotent stem cell-derived kidney organoids for personalized congenital and idiopathic nephrotic syndrome modeling

10.1101/2021.10.27.466054 ◽

2021 ◽

Author(s):

Jitske Jansen ◽

Bartholomeus T van den Berge ◽

Martijn van den Broek ◽

Rutger J Maas ◽

Brigith Willemsen ◽

...

Keyword(s):

Nephrotic Syndrome ◽

Stem Cell ◽

Pluripotent Stem Cell ◽

Idiopathic Nephrotic Syndrome ◽

Super Resolution ◽

Induced Pluripotent Stem Cell ◽

Glomerular Injury ◽

Kidney Organoids

Nephrotic syndrome (NS) is characterized by severe proteinuria as a consequence of kidney glomerular injury due to podocyte damage. In vitro models mimicking in vivo podocyte characteristics are a prerequisite to resolve NS pathogenesis. Here, we report human induced pluripotent stem cell derived kidney organoids containing a podocyte population that heads towards adult podocytes and were superior compared to 2D counterparts, based on scRNA sequencing, super-resolution imaging and electron microscopy. In this study, these next-generation podocytes in kidney organoids enabled personalized idiopathic nephrotic syndrome modeling as shown by activated slit diaphragm signaling and podocyte injury following protamine sulfate treatment and exposure to NS plasma containing pathogenic permeability factors. Organoids cultured from cells of a patient with heterozygous NPHS2 mutations showed poor NPHS2 expression and aberrant NPHS1 localization, which was reversible after genetic correction. Repaired organoids displayed increased VEGFA pathway activity and transcription factor activity known to be essential for podocyte physiology, as shown by RNA sequencing. This study shows that organoids are the preferred model of choice to study idiopathic and congenital podocytopathies.

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20 In vivo tracking of human pluripotent stem cell vascular derivatives.

Heart ◽

10.1136/heartjnl-2012-302951.020 ◽

2012 ◽

Vol 98 (Suppl 3) ◽

pp. A6.3-A6

Author(s):

L Low ◽

C Cheung ◽

M Bennett ◽

S Sinha

Keyword(s):

Stem Cell ◽

Pluripotent Stem Cell ◽

Human Pluripotent Stem Cell

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Pluripotent stem cell-derived kidney organoids: An in vivo-like in vitro technology

European Journal of Pharmacology ◽

10.1016/j.ejphar.2016.06.059 ◽

2016 ◽

Vol 790 ◽

pp. 12-20 ◽

Cited By ~ 20

Author(s):

Frans Schutgens ◽

Marianne C. Verhaar ◽

Maarten B. Rookmaaker

Keyword(s):

Stem Cell ◽

Pluripotent Stem Cell ◽

Kidney Organoids

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Close neighbors in the niche: Paracrine signals from endothelial cells promote alveolar epithelial differentiation of induced pluripotent stem cell-derived lung progenitors

Cytotherapy ◽

10.1016/j.jcyt.2018.02.045 ◽

2018 ◽

Vol 20 (5) ◽

pp. S21

Author(s):

M. Ho ◽

M.S. Ho ◽

D.J. Stewart

Keyword(s):

Endothelial Cells ◽

Stem Cell ◽

Pluripotent Stem Cell ◽

Induced Pluripotent Stem Cell ◽

Epithelial Differentiation ◽

Alveolar Epithelial ◽

Lung Progenitors ◽

Induced Pluripotent

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Regulation of JAM2 Expression in the Lungs of Streptozotocin-Induced Diabetic Mice and Human Pluripotent Stem Cell-Derived Alveolar Organoids

Biomedicines ◽

10.3390/biomedicines8090346 ◽

2020 ◽

Vol 8 (9) ◽

pp. 346 ◽

Cited By ~ 2

Author(s):

Roya Rasaei ◽

Eunbi Kim ◽

Ji-Young Kim ◽

Sunghun Na ◽

Jung-Hyun Kim ◽

...

Keyword(s):

Stem Cell ◽

Pluripotent Stem Cell ◽

Causative Factor ◽

Human Mesenchymal Stem Cell ◽

Therapeutic Effects ◽

Diabetic Mice ◽

Human Pluripotent Stem Cell ◽

Potential Indicator

Hyperglycemia is a causative factor in the pathogenesis of respiratory diseases, known to induce fibrosis and inflammation in the lung. However, little attention has been paid to genes related to hyperglycemic-induced lung alterations and stem cell applications for therapeutic use. In this study, our microarray data revealed significantly increased levels of junctional adhesion molecule 2 (JAM2) in the high glucose (HG)-induced transcriptional profile in human perivascular cells (hPVCs). The elevated level of JAM2 in HG-treated hPVCs was transcriptionally and epigenetically reversible when HG treatment was removed. We further investigated the expression of JAM2 using in vivo and in vitro hyperglycemic models. Our results showed significant upregulation of JAM2 in the lungs of streptozotocin (STZ)-induced diabetic mice, which was greatly suppressed by the administration of conditioned medium obtained from human mesenchymal stem cell cultures. Furthermore, JAM2 was found to be significantly upregulated in human pluripotent stem cell-derived multicellular alveolar organoids by exposure to HG. Our results suggest that JAM2 may play an important role in STZ-induced lung alterations and could be a potential indicator for predicting the therapeutic effects of stem cells and drugs in diabetic lung complications.

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