Pluripotent Stem Cells

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13 pages, 5125 KiB

Open AccessArticle

Pilot Study for Isolation of Stromal Vascular Fraction with Collagenase Using an Automated Processing System

by Gershon Zinger, Yoav Gronovich, Adi Maisel Lotan and Racheli Sharon-Gabbay

Int. J. Mol. Sci. 2024, 25(13), 7148; https://doi.org/10.3390/ijms25137148 - 28 Jun 2024

Viewed by 761

Abstract

There are many potential therapeutic applications for autologous adipose-derived stromal cells. These cells are found in a heterogeneous population isolated from adipose tissue called the stromal vascular fraction (SVF). Closed automated systems are available to release cells from the adherent stroma. Here, we test one system to evaluate the heterogeneous output for yield, purity, cellular characterization, and stemness criteria. The SVF was isolated from three donors using the Automated Cell Station (ACS) from BSL Co., Ltd., Busan, Republic of Korea. The SVF cellular output was characterized for cell yield and viability, immunophenotyping analysis, pluripotent differentiation potential, adhesion to plastic, and colony-forming units. Additionally, the SVF was tested for endotoxin and collagenase residuals. The SVF yield from the ACS system was an average volume of 7.9 ± 0.5 mL containing an average of 19 × 10⁶ nucleated cells with 85 ± 12% viability. Flow cytometry identified a variety of cells, including ASCs (23%), macrophages (24%), endothelial cells (5%), pericytes (4%), and transitional cells (0.5%). The final concentrated product contained cells capable of differentiating into adipogenic, chondrogenic, and osteogenic phenotypes. Furthermore, tests for SVF sterility and purity showed no evidence of endotoxin or collagenase residuals. The ACS system can efficiently process cells from adipose tissue within the timeframe of a single surgical procedure. The cellular characterization indicated that this system can yield a sterile and concentrated SVF output, providing a valuable source of ASCs within the heterogeneous cell population. Full article

(This article belongs to the Topic Pluripotent Stem Cells)

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24 pages, 9421 KiB

Open AccessArticle

MiR-290 Family Maintains Pluripotency and Self-Renewal by Regulating MAPK Signaling Pathway in Intermediate Pluripotent Stem Cells

by Yueshi Liu, Xiangnan Li, Xiaozhuang Ma, Qiankun Du, Jiemin Wang and Haiquan Yu

Int. J. Mol. Sci. 2024, 25(5), 2681; https://doi.org/10.3390/ijms25052681 - 26 Feb 2024

Viewed by 1384

Abstract

Mouse embryonic stem cells (ESCs) and epiblast stem cells (EpiSCs) are derived from pre- and post-implantation embryos, representing the initial “naïve” and final “primed” states of pluripotency, respectively. In this study, novel reprogrammed pluripotent stem cells (rPSCs) were induced from mouse EpiSCs using a chemically defined medium containing mouse LIF, BMP4, CHIR99021, XAV939, and SB203580. The rPSCs exhibited domed clones and expressed key pluripotency genes, with both X chromosomes active in female cells. Furthermore, rPSCs differentiated into cells of all three germ layers in vivo through teratoma formation. Regarding epigenetic modifications, the DNA methylation of Oct4, Sox2, and Nanog promoter regions and the mRNA levels of Dnmt3a, Dnmt3b, and Dnmt1 were reduced in rPSCs compared with EpiSCs. However, the miR-290 family was significantly upregulated in rPSCs. After removing SB203580, an inhibitor of the p38 MAPK pathway, the cell colonies changed from domed to flat, with a significant decrease in the expression of pluripotency genes and the miR-290 family. Conversely, overexpression of pri-miR-290 reversed these changes. In addition, Map2k6 was identified as a direct target gene of miR-291b-3p, indicating that the miR-290 family maintains pluripotency and self-renewal in rPSCs by regulating the MAPK signaling pathway. Full article

(This article belongs to the Topic Pluripotent Stem Cells)

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17 pages, 2214 KiB

Open AccessPerspective

The Management of Data for the Banking, Qualification, and Distribution of Induced Pluripotent Stem Cells: Lessons Learned from the European Bank for Induced Pluripotent Stem Cells

by Nancy Mah, Andreas Kurtz, Antonie Fuhr, Stefanie Seltmann, Ying Chen, Nils Bultjer, Johannes Dewender, Ayuen Lual, Rachel Steeg and Sabine C. Mueller

Cells 2023, 12(23), 2756; https://doi.org/10.3390/cells12232756 - 1 Dec 2023

Cited by 2 | Viewed by 1344

Abstract

The European Bank for induced pluripotent Stem Cells (EBiSC) was established in 2014 as a non-profit project for the banking, quality control, and distribution of human iPSC lines for research around the world. EBiSC iPSCs are deposited from diverse laboratories internationally and, hence, a key activity for EBiSC is standardising not only the iPSC lines themselves but also the data associated with them. This includes enabling unique nomenclature for the cells, as well as applying uniformity to the data provided by the cell line generator versus quality control data generated by EBiSC, and providing mechanisms to share personal data in a secure and GDPR-compliant manner. A joint approach implemented by EBiSC and the human pluripotent stem cell registry (hPSCreg^®) has provided a solution that enabled hPSCreg^® to improve its registration platform for iPSCs and EBiSC to have a pipeline for the import, standardisation, storage, and management of data associated with EBiSC iPSCs. In this work, we describe the experience of cell line data management for iPSC banking throughout the course of EBiSC’s development as a central European banking infrastructure and present a model for how this could be implemented by other iPSC repositories to increase the FAIRness of iPSC research globally. Full article

(This article belongs to the Topic Pluripotent Stem Cells)

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12 pages, 1536 KiB

Open AccessArticle

MPP8 Governs the Activity of the LIF/STAT3 Pathway and Plays a Crucial Role in the Differentiation of Mouse Embryonic Stem Cells

by Heyao Zhang, Tenghui Yang, Hao Wu, Wen Yi, Chunhong Dai, Xi Chen, Wensheng Zhang and Ying Ye

Cells 2023, 12(16), 2023; https://doi.org/10.3390/cells12162023 - 8 Aug 2023

Cited by 1 | Viewed by 1958

Abstract

Mouse embryonic stem cells (mESCs) possess the remarkable characteristics of unlimited self-renewal and pluripotency, which render them highly valuable for both fundamental research and clinical applications. A comprehensive understanding of the molecular mechanisms underlying mESC function is of the utmost importance. The Human Silence Hub (HUSH) complex, comprising FAM208A, MPP8, and periphilin, constitutes an epigenetic silencing complex involved in suppressing retroviruses and transposons during early embryonic development. However, its precise role in regulating mESC pluripotency and differentiation remains elusive. In this study, we generated homogenous miniIAA7-tagged Mpp8 mouse ES cell lines. Upon induction of MPP8 protein degradation, we observed the impaired proliferation and reduced colony formation ability of mESCs. Furthermore, this study unveils the involvement of MPP8 in regulating the activity of the LIF/STAT3 signaling pathway and Nanog expression in mESCs. Finally, we provide compelling evidence that degradation of the MPP8 protein impairs the differentiation of mESC. Full article

(This article belongs to the Topic Pluripotent Stem Cells)

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22 pages, 6893 KiB

Open AccessArticle

Mixed-Lineage Leukemia 1 Inhibition Enhances the Differentiation Potential of Bovine Embryonic Stem Cells by Increasing H3K4 Mono-Methylation at Active Promoters

by Chen Li, Xuejie Han, Jing Wang, Fang Liu, Yuanyuan Zhang, Zihong Li, Zhenyu Lu, Yongli Yue, Jinzhu Xiang and Xueling Li

Int. J. Mol. Sci. 2023, 24(15), 11901; https://doi.org/10.3390/ijms241511901 - 25 Jul 2023

Viewed by 1501

Abstract

Mixed-lineage leukemia 1 (MLL1) introduces 1-, 2- and 3-methylation into histone H3K4 through the evolutionarily conserved set domain. In this study, bovine embryonic stem cells (bESCs, known as bESCs-F7) were established from in vitro-fertilized (IVF) embryos via Wnt signaling inhibition; however, their contribution to the endoderm in vivo is limited. To improve the quality of bESCs, MM-102, an inhibitor of MLL1, was applied to the culture. The results showed that MLL1 inhibition along with GSK3 and MAP2K inhibition (3i) at the embryonic stage did not affect bESCs’ establishment and pluripotency. MLL1 inhibition improved the pluripotency and differentiation potential of bESCs via the up-regulation of stem cell signaling pathways such as PI3K-Akt and WNT. MLL1 inhibition decreased H3K4me1 modification at the promoters and altered the distribution of DNA methylation in bESCs. In summary, MLL1 inhibition gives bESCs better pluripotency, and its application may provide high-quality pluripotent stem cells for domestic animals. Full article

(This article belongs to the Topic Pluripotent Stem Cells)

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18 pages, 4084 KiB

Open AccessArticle

Cerebral-Organoid-Derived Exosomes Alleviate Oxidative Stress and Promote LMX1A-Dependent Dopaminergic Differentiation

by Xingrui Ji, Shaocong Zhou, Nana Wang, Jingwen Wang, Yue Wu, Yuhan Duan, Penghao Ni, Jingzhong Zhang and Shuang Yu

Int. J. Mol. Sci. 2023, 24(13), 11048; https://doi.org/10.3390/ijms241311048 - 4 Jul 2023

Cited by 5 | Viewed by 2824

Abstract

The remarkable advancements related to cerebral organoids have provided unprecedented opportunities to model human brain development and diseases. However, despite their potential significance in neurodegenerative diseases such as Parkinson’s disease (PD), the role of exosomes from cerebral organoids (OExo) has been largely unknown. In this study, we compared the effects of OExo to those of mesenchymal stem cell (MSC)-derived exosomes (CExo) and found that OExo shared similar neuroprotective effects to CExo. Our findings showed that OExo mitigated H₂O₂-induced oxidative stress and apoptosis in rat midbrain astrocytes by reducing excess ROS production, antioxidant depletion, lipid peroxidation, mitochondrial dysfunction, and the expression of pro-apoptotic genes. Notably, OExo demonstrated superiority over CExo in promoting the differentiation of human-induced pluripotent stem cells (iPSCs) into dopaminergic (DA) neurons. This was attributed to the higher abundance of neurotrophic factors, including neurotrophin-4 (NT-4) and glial-cell-derived neurotrophic factor (GDNF), in OExo, which facilitated the iPSCs’ differentiation into DA neurons in an LIM homeobox transcription factor 1 alpha (LMX1A)-dependent manner. Our study provides novel insight into the biological properties of cerebral organoids and highlights the potential of OExo in the treatment of neurodegenerative diseases such as PD. Full article

(This article belongs to the Topic Pluripotent Stem Cells)

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20 pages, 9868 KiB

Open AccessArticle

Essential Role of Adhesion GPCR, GPR123, for Human Pluripotent Stem Cells and Reprogramming towards Pluripotency

by Olga A. Krasnova, Karina A. Kulakova, Julia V. Sopova, Evgenyi Y. Smirnov, Sergey A. Silonov, Ekaterina V. Lomert, Olga A. Bystrova, Marina G. Martynova and Irina E. Neganova

Cells 2023, 12(2), 304; https://doi.org/10.3390/cells12020304 - 13 Jan 2023

Cited by 1 | Viewed by 2407

Abstract

G-protein-coupled receptors (GPCRs) are the largest family of cell surface receptors. They modulate key physiological functions and are required in diverse developmental processes including embryogenesis, but their role in pluripotency maintenance and acquisition during the reprogramming towards hiPSCs draws little attention. Meanwhile, it is known that more than 106 GPCRs are overexpressed in human pluripotent stem cells (hPSCs). Previously, to identify novel effectors of reprogramming, we performed a high-throughput RNA interference (RNAi) screening assay and identified adhesion GPCR, GPR123, as a potential reprogramming effector. Its role has not been explored before. Herein, by employing GPR123 RNAi we addressed the role of GPR123 for hPSCs. The suppression of GPR123 in hPSCs leads to the loss of pluripotency and differentiation, impacted colony morphology, accumulation of cells at the G2 phase of the cell cycle, and absence of the scratch closure. Application of the GPR123 RNAi at the initiation stage of reprogramming leads to a decrease in the percentage of the “true” hiPSC colonies, a drop in E-cadherin expression, a decrease in the percentage of NANOG+ nuclei, and the absence of actin cytoskeleton remodeling. Together this leads to the absence of the alkaline-phosphatase-positive hiPSCs colonies on the 18th day of the reprogramming process. Overall, these data indicate for the first time the essential role of GPR123 in the maintenance and acquisition of pluripotency. Full article

(This article belongs to the Topic Pluripotent Stem Cells)

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12 pages, 8556 KiB

Open AccessBrief Report

Bcl-xL Promotes the Survival of Motor Neurons Derived from Neural Stem Cells

by Yunqin Wu, Xiaohua Peng, Song Ang, Yue Gao, Yue Chi, Jinling Wang, Chengcheng Tang, Xiaoqing Zhou, Yanxian Feng, Kun Zhang, Qingjian Zou and Min Chen

Biology 2023, 12(1), 132; https://doi.org/10.3390/biology12010132 - 13 Jan 2023

Cited by 1 | Viewed by 1967

Abstract

Neural stem cell (NSC) transplantation creates new hope for the treatment of neurodegenerative disorders by direct differentiation into neurons. However, this technique is limited by poor survival and functional neuron deficiency. In this research study, we generated pro-survival murine NSCs (mNSCs) via the ectopic expression of Bcl-xL. A doxycycline (Dox)-inducible Ngn2-Isl1-Lhx3 system was also integrated into the mNSC genome. The four gene-modified mNSCs can rapidly and effectively differentiate into motor neurons after Dox treatments. Ectopic Bcl-xL could resist replating-induced stress, glutamate toxicity, neuronal apoptosis and remarkably promote the survival of motor neurons. Taken together, we established genetically modified mNSCs with improved survival, which may be useful for motor neuron degenerative diseases. Full article

(This article belongs to the Topic Pluripotent Stem Cells)

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Journal Name	Impact Factor	CiteScore	Launched Year	First Decision (median)	APC
Biology biology	3.6	5.7	2012	16.1 Days	CHF 2700
Cells cells	5.1	9.9	2012	17.5 Days	CHF 2700
International Journal of Molecular Sciences ijms	4.9	8.1	2000	18.1 Days	CHF 2900
Organoids organoids	-	-	2022	15.0 days *	CHF 1000

* Median value for all MDPI journals in the first half of 2024.

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