scholarly journals FTIR Spectroscopic and Molecular Analysis during Differentiation of Pluripotent Stem Cells to Pancreatic Cells

2016 ◽  
Vol 2016 ◽  
pp. 1-10 ◽  
Author(s):  
Gustavo Jesus Vazquez-Zapien ◽  
Monica Maribel Mata-Miranda ◽  
Virginia Sanchez-Monroy ◽  
Raul Jacobo Delgado-Macuil ◽  
David Guillermo Perez-Ishiwara ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Ftir Spectra ◽  
Pancreas Development ◽  
Differentiation Process ◽  
Absorption Bands ◽  
Biophysical Parameter ◽  
Real Time Quantitative Pcr ◽  
Pancreatic Cells ◽  
Embryonic Pancreas

Some of the greatest challenges in stem cells (SCs) biology and regenerative medicine are differentiation control of SCs and ensuring the purity of differentiated cells. In this work, we differentiated mouse pluripotent stem cells (mPSCs) toward pancreatic cells characterizing this differentiation process by molecular and spectroscopic technics. Both mPSCs and Differentiated Pancreatic Cells (DPCs) were subjected to a genetic, phenotypic, and biochemical analysis by real-time quantitative PCR (RT-qPCR), immunocytochemistry, and Fourier Transform Infrared (FTIR) spectroscopy. Cultured mPCSs expressed pluripotent genes and proteins (NanogandSOX2). DPCs expressed endodermal genes (SOX17andPdx1) at day 11, an inductor gene of embryonic pancreas development (Pdx1) at day 17 and pancreas genes and proteins (InsulinandGlucagon) at day 21 of differentiation. Likewise, FTIR spectra of mPSCs and DPCs at different maturation stages (11, 17, and 21 days) were obtained and showed absorption bands related with different types of biomolecules. These FTIR spectra exhibited significant spectral changes agreeing with the differentiation process, particularly in proteins and nucleic acids bands. In conclusion, the obtained DPCs passed through the chronological stages of embryonic pancreas development and FTIR spectra provide a new biophysical parameter based on molecular markers indicating the differentiation process of mPSCs to specialized cells.

scholarly journals Deconstructing Pancreas Development to Reconstruct Human Islets from Pluripotent Stem Cells

2010 ◽  
Vol 6 (4) ◽  
pp. 300-308 ◽  
Author(s):  
Kristen D. McKnight ◽  
Pei Wang ◽  
Seung K. Kim
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Human Islets ◽  
Pancreas Development

scholarly journals Establish deeper understanding on the osteogenic differentiation of monolayer cultured human pluripotent stem cells using novel and detail analyses

2020 ◽  
Author(s):  
Ping Zhou ◽  
Jia-Min Shi ◽  
Jing-E Song ◽  
Yu Han ◽  
Hong-Jiao Li ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Pluripotent Stem Cells ◽  
Telomerase Activity ◽  
Human Pluripotent Stem Cells ◽  
Cell Counting ◽  
Integrated Analysis ◽  
Differentiation Process ◽  
Cell Type

Abstract Background: Derivation of the osteoblast-like cell from human pluripotent stem cell becomes a hot topic in bone tissue engineering. Although many improvements have been achieved in this field, low induction efficiency because of the non-directed differentiation process hampers their application in bone regeneration. We think lack of detailed understanding on the osteogenic differentiation process should be the main reason.Methods: Monolayer cultured human embryonic stem cells and human induced pluripotent stem cells were inducted in traditional serum-containing osteogenic medium for 35 days. Except for traditional assays such as cell viability detection, reverse transcription-polymerase chain reaction, immunofluorescence, and alizarin red staining, we also applied cell counting, cell telomerase activity, cell cycle and quantitative expression of runt-related transcription factor 2 as essential indicators to analyze the cell type changes during the differentiation process. Results: The population of differentiated cells are quite heterogenous throughout 35 days of induction. Then, cell telomerase activity and cell cycle analyses have value in evaluating the cell type changes and tumorigenicity of obtained cells. Moreover, nuclear staining should be a recommended method to evaluate the cell number, because, it is still a great challenge to dissociate cells with varying differentiation times into single cells with high survival rate. Finally, a dynamic map was made to integrated analysis of these results, and the cell types at defined stages of osteogenic differentiation of human pluripotent stem cells was concluded. Conclusions: This study lay foundation to improve the in vitro osteogenic differentiation efficiency of human pluripotent stem cells by supplementing functional compounds at each stage according to a time-frame, then establish a step-wised induction system in the future.

scholarly journals Principal Components by FTIR Spectroscopy as Innovative Characterization Technique during Differentiation of Pluripotent Stem Cells to Pancreatic Cells

2017 ◽  
Author(s):  
◽  
M. M. Mata-Miranda
Keyword(s):  
Stem Cells ◽  
Ftir Spectroscopy ◽  
Pluripotent Stem Cells ◽  
Principal Component ◽  
Cell Lineages ◽  
Non Invasive ◽  
Differentiated Cells ◽  
Pancreatic Cells ◽  
Second Derivatives

Two of the greatest challenges in Stem Cells (SCs) biology and regenerative medicine, are differentiation control of SCs and ensuring the purity of differentiated cells. In this sense, fast, efficient and accurate techniques for SCs characterization and their differentiation into different cell lineages are needed. The aim of this study was to analyse Pluripotent Stem Cells (PSCs) and Differentiated Pancreatic Cells (DPCs) by Fourier Transform Infrared (FTIR) spectroscopy and Principal Component Analysis (PCA). For this purpose, we differentiated PSCs toward DPCs, characterizing the differentiation process at different stages (0, 11, 17 and 21 days) through light microscopy and vibrational spectroscopy. FTIR spectra were analysed with the multivariate method of PCA, using the second derivatives in the protein, carbohydrate and ribose regions. The results indicate that the PCA allows to characterize and discriminate PSCs and DPCs at different stages of differentiation in the analysed spectral regions. From these results, we concluded that the PCA allows the chemically and structural characterization of PSCs and the different stages of their differentiation in a fast, accurate and non-invasive way.

scholarly journals BCL-xL/BCL2L1 is a critical anti-apoptotic protein that promotes the survival of differentiating pancreatic cells from human pluripotent stem cells

2020 ◽  
Vol 11 (5) ◽  
Author(s):  
Larry Sai Weng Loo ◽  
Andreas Alvin Purnomo Soetedjo ◽  
Hwee Hui Lau ◽  
Natasha Hui Jin Ng ◽  
Soumita Ghosh ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Pancreatic Cells ◽  
Apoptotic Protein

scholarly journals HERVH-derived lncRNAs negatively regulate chromatin targeting and remodeling mediated by CHD7

2021 ◽  
Vol 5 (1) ◽  
pp. e202101127
Author(s):  
Fu-Kai Hsieh ◽  
Fei Ji ◽  
Manashree Damle ◽  
Ruslan I Sadreyev ◽  
Robert E Kingston
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
De Novo ◽  
Charge Syndrome ◽  
Differentiation Process ◽  
De Novo Mutations ◽  
Pluripotent Cells ◽  
Developmental Defects ◽  
High Affinity ◽  
And Function

Chd7 encodes an ATP-dependent chromatin remodeler which has been shown to target specific genomic loci and alter local transcription potentially by remodeling chromatin structure. De novo mutations in CHD7 are the major cause of CHARGE syndrome which features multiple developmental defects. We examined whether nuclear RNAs might contribute to its targeting and function and identified a preferential interaction between CHD7 and lncRNAs derived from HERVH loci in pluripotent stem cells. Knockdown of HERVH family lncRNAs using LNAs or knockout of an individual copy of HERVH by CRISPR-Cas9 both resulted in increased binding of CHD7 and increased levels of H3K27ac at a subset of enhancers. Depletion of HERVH family RNAs led to the activation of multiple genes. CHD7 bound HERVH RNA with high affinity but low specificity and this interaction decreased the ability of CHD7 to bind and remodel nucleosomes. We present a model in which HERVH lncRNAs act as a decoy to modulate the dynamics of CHD7 binding to enhancers in pluripotent cells and the activation of numerous genes that might impact the differentiation process.

scholarly journals Modeling Maturity Onset Diabetes of the Young in Pluripotent Stem Cells: Challenges and Achievements

2021 ◽  
Vol 12 ◽  
Author(s):  
Carmel Braverman-Gross ◽  
Nissim Benvenisty
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Monogenic Diabetes ◽  
Human Cells ◽  
Rodent Models ◽  
Maturity Onset Diabetes ◽  
Great Opportunity ◽  
Pancreatic Cells ◽  
Onset Diabetes

Maturity onset diabetes of the young (MODY), is a group of monogenic diabetes disorders. Rodent models for MODY do not fully recapitulate the human phenotypes, calling for models generated in human cells. Human pluripotent stem cells (hPSCs), capable of differentiation towards pancreatic cells, possess a great opportunity to model MODY disorders in vitro. Here, we review the models for MODY diseases in hPSCs to date and the molecular lessons learnt from them. We also discuss the limitations and challenges that these types of models are still facing.

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