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 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 DNA damage and repair in differentiation of stem cells and cells of connective cell lineages: A trigger or a complication?

2021 ◽  
Vol 65 (2) ◽  
Author(s):  
Nikolajs Sjakste ◽  
Una Riekstiņa
Keyword(s):  
Stem Cells ◽  
Dna Damage ◽  
Dna Repair ◽  
Pluripotent Stem Cells ◽  
Differentiation Process ◽  
Dna Breaks ◽  
Repair Capacity ◽  
Cell Lineages ◽  
Dna Repair Capacity ◽  
Differentiated Cells

The review summarizes literature data on the role of DNA breaks and DNA repair in differentiation of pluripotent stem cells (PSC) and connective cell lineages. PSC, including embryonic stem cells (ESC) and induced pluripotent stem cells (iPSC), are rapidly dividing cells with highly active DNA damage response (DDR) mechanisms to ensure the stability and integrity of the DNA. In PSCs, the most common DDR mechanism is error-free homologous recombination (HR) that is primarily active during S phase of the cell cycle, whereas in quiescent, slow-dividing or non-dividing tissue progenitors and terminally differentiated cells, error-prone non-homologous end joining (NHEJ) mechanism of the double-strand break (DSB) repair is dominating.  Thus, it seems that reprogramming and differentiation induce DNA strand breaks in stem cells which itself may trigger the differentiation process. Somatic cell reprogramming to iPSCs is preceded by a transient increase of the DSBs induced presumably by the caspase-dependent DNase or reactive oxygen species (ROS). In general, pluripotent stem cells possess stronger DNA repair systems compared to the differentiated cells. Nonetheless, during a prolonged cell culture propagation, DNA breaks can accumulate due to the DNA polymerase stalling. Consequently, the DNA damage might trigger the differentiation of stem cells or a replicative senescence of somatic cells. Differentiation process per se is often accompanied by a decrease of the DNA repair capacity. Thus, the differentiation might be triggered by DNA breaks, alternatively the breaks can be a consequence of the decay in the DNA repair capacity of differentiated cells.

Derivation and Characterization of human Induced Pluripotent Stem Cells

Acta Naturae ◽  
2009 ◽  
Vol 1 (2) ◽  
pp. 91-92 ◽  
Author(s):  
M V Shutova ◽  
A N Bogomazova ◽  
M A Lagarkova ◽  
S L Kiselev
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Induced Pluripotent

Mini Review; Differentiation of Human Pluripotent Stem Cells into Oocytes

2020 ◽  
Vol 15 (4) ◽  
pp. 301-307 ◽  
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.

scholarly journals A non-invasive method to generate induced pluripotent stem cells from primate urine

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Johanna Geuder ◽  
Lucas E. Wange ◽  
Aleksandar Janjic ◽  
Jessica Radmer ◽  
Philipp Janssen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Sendai Virus ◽  
Cell Types ◽  
Urine Samples ◽  
Comparative Approach ◽  
Non Invasive ◽  
Human Ipscs ◽  
Induced Pluripotent

AbstractComparing the molecular and cellular properties among primates is crucial to better understand human evolution and biology. However, it is difficult or ethically impossible to collect matched tissues from many primates, especially during development. An alternative is to model different cell types and their development using induced pluripotent stem cells (iPSCs). These can be generated from many tissue sources, but non-invasive sampling would decisively broaden the spectrum of non-human primates that can be investigated. Here, we report the generation of primate iPSCs from urine samples. We first validate and optimize the procedure using human urine samples and show that suspension- Sendai Virus transduction of reprogramming factors into urinary cells efficiently generates integration-free iPSCs, which maintain their pluripotency under feeder-free culture conditions. We demonstrate that this method is also applicable to gorilla and orangutan urinary cells isolated from a non-sterile zoo floor. We characterize the urinary cells, iPSCs and derived neural progenitor cells using karyotyping, immunohistochemistry, differentiation assays and RNA-sequencing. We show that the urine-derived human iPSCs are indistinguishable from well characterized PBMC-derived human iPSCs and that the gorilla and orangutan iPSCs are well comparable to the human iPSCs. In summary, this study introduces a novel and efficient approach to non-invasively generate iPSCs from primate urine. This will extend the zoo of species available for a comparative approach to molecular and cellular phenotypes.

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

Cells ◽  
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.

scholarly journals Generation and characterization of human induced pluripotent stem cells (iPSCs) from three male and three female patients with CDKL5 Deficiency Disorder (CDD)

2021 ◽  
Vol 53 ◽  
pp. 102276
Author(s):  
Pin-Fang Chen ◽  
Teresa Chen ◽  
Taylor E. Forman ◽  
Amanda C. Swanson ◽  
Benjamin O'Kelly ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Female Patients ◽  
Induced Pluripotent

scholarly journals Physical cues of cell culture materials lead the direction of differentiation lineages of pluripotent stem cells

2015 ◽  
Vol 3 (41) ◽  
pp. 8032-8058 ◽  
Author(s):  
Akon Higuchi ◽  
Qing-Dong Ling ◽  
S. Suresh Kumar ◽  
Yung Chang ◽  
Abdullah A. Alarfaj ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Pluripotent Stem Cells ◽  
Specific Cell ◽  
Cell Lineages ◽  
Physical Cues

Differentiation methods of hPSCs into specific cell lineages. Differentiation of hPSCsviaEB formation (types AB, A–D) or without EB formation (types E–H).

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