Turning Potential Into Action: Using Pluripotent Stem Cells to Understand Heart Development and Function in Health and Disease

AbstractSimultaneous differentiation of human induced pluripotent stem cells (hiPSCs) into divergent cell types offers a pathway to achieving tailorable cellular complexity, patterned architecture, and function in engineered human organoids and tissues. Recent transcription factor (TF) overexpression protocols typically produce only one cell type of interest rather than the multitude of cell types and structural organization found in native human tissues. Here, we report an orthogonal differentiation platform for genomically programming stem cells, organoids and bioprinted tissues with controlled composition and organization. To demonstrate this platform, we orthogonally differentiated endothelial cells and neurons from hiPSCs in a one-pot system containing neural stem cell-specifying media. By aggregating inducible-TF and wildtype hiPSCs into pooled and multicore-shell embryoid bodies, we produced vascularized and patterned cortical organoids within days. Using multimaterial 3D bioprinting, we patterned 3D neural tissues from densely cellular, matrix-free stem cell inks that were orthogonally differentiated on demand into distinct layered regions composed of neural stem cells, endothelium, and neurons, respectively. Given the high proliferative capacity and patient-specificity of hiPSCs, our platform provides a facile route for programming cells and multicellular tissues for drug screening and therapeutic applications.

Download Full-text

Analysis of the expression of PIWI-interacting RNAs during cardiac differentiation of human pluripotent stem cells

10.1101/639906 ◽

2019 ◽

Author(s):

Alejandro La Greca ◽

María Agustina Scarafía ◽

María Clara Hernández Cañás ◽

Nelba Pérez ◽

Sheila Castañeda ◽

...

Keyword(s):

Stem Cells ◽

Pluripotent Stem Cells ◽

Muscle Development ◽

Germ Line ◽

Cardiac Cells ◽

Cardiac Differentiation ◽

Fine Tuning ◽

Protein Coding ◽

Sense Strand ◽

And Function

SummaryPIWI-interacting RNAs (piRNAs) are a class of non-coding RNAs initially thought to be restricted almost exclusively to germ line cells. In recent years, accumulating evidence has demonstrated that piRNAs are actually expressed in somatic cells like pluripotent, neural, cardiac and even cancer cells. However, controversy still remains around the existence and function of somatic piRNAs. Using small RNA-seq samples from H9 pluripotent stem cells differentiated to mesoderm progenitors and cardiomyocytes we identified the expression of 447 piRNAs, of which 241 were detected in pluripotency, 218 in mesoderm and 171 in cardiac cells. The majority of them originated from the sense strand of protein coding and lncRNAs genes in all stages of differentiation, though no evidences for secondary piRNAs (ping-pong loop) were found. Genes hosting piRNAs in cardiac samples were related to critical biological processes in the heart, like contraction and cardiac muscle development. Our results indicate that somatic piRNAs might have a role in fine-tuning the expression of genes involved in the differentiation of pluripotent cells to cardiomyocytes.

Download Full-text

Efficient derivation of human trophoblast stem cells from primed pluripotent stem cells

Science Advances ◽

10.1126/sciadv.abf4416 ◽

2021 ◽

Vol 7 (33) ◽

pp. eabf4416

Author(s):

Yanxing Wei ◽

Tianyu Wang ◽

Lishi Ma ◽

Yanqi Zhang ◽

Yuan Zhao ◽

...

Keyword(s):

Stem Cells ◽

Pluripotent Stem Cells ◽

Chromatin Accessibility ◽

Placental Development ◽

Differentiation Potential ◽

Human Trophoblast ◽

Trophoblast Stem Cells ◽

Trophoblast Stem ◽

Early Placenta ◽

And Function

Human trophoblast stem cells (hTSCs) provide a valuable model to study placental development and function. While primary hTSCs have been derived from embryos/early placenta, and transdifferentiated hTSCs from naïve human pluripotent stem cells (hPSCs), the generation of hTSCs from primed PSCs is problematic. We report the successful generation of TSCs from primed hPSCs and show that BMP4 substantially enhances this process. TSCs derived from primed hPSCs are similar to blastocyst-derived hTSCs in terms of morphology, proliferation, differentiation potential, and gene expression. We define the chromatin accessibility dynamics and histone modifications (H3K4me3/H3K27me3) that specify hPSC-derived TSCs. Consistent with low density of H3K27me3 in primed hPSC-derived hTSCs, we show that knockout of H3K27 methyltransferases (EZH1/2) increases the efficiency of hTSC derivation from primed hPSCs. Efficient derivation of hTSCs from primed hPSCs provides a simple and powerful model to understand human trophoblast development, including the pathogenesis of trophoblast-related disorders, by generating disease-specific hTSCs.

Download Full-text

Human Pluripotent Stem Cells to Model Islet Defects in Diabetes

Frontiers in Endocrinology ◽

10.3389/fendo.2021.642152 ◽

2021 ◽

Vol 12 ◽

Author(s):

Diego Balboa ◽

Diepiriye G. Iworima ◽

Timothy J. Kieffer

Keyword(s):

Stem Cells ◽

Beta Cells ◽

Pluripotent Stem Cells ◽

Pancreatic Beta Cells ◽

Molecular Mechanisms ◽

Disease Modeling ◽

Human Pluripotent Stem Cells ◽

Pancreatic Endocrine Cells ◽

Onset Of Diabetes ◽

And Function

Diabetes mellitus is characterized by elevated levels of blood glucose and is ultimately caused by insufficient insulin production from pancreatic beta cells. Different research models have been utilized to unravel the molecular mechanisms leading to the onset of diabetes. The generation of pancreatic endocrine cells from human pluripotent stem cells constitutes an approach to study genetic defects leading to impaired beta cell development and function. Here, we review the recent progress in generating and characterizing functional stem cell-derived beta cells. We summarize the diabetes disease modeling possibilities that stem cells offer and the challenges that lie ahead to further improve these models.

Download Full-text