scholarly journals Reprogramming By Cytosolic Extract of Human Embryonic Stem Cells Improves Dopaminergic Differentiation Potential of Human Adipose Tissue-Derived Stem Cells

2021 ◽  
Author(s):  
Sehar Mobasseri ◽  
◽  
Arash Javeri ◽  
Masoumeh Fakhr Taha ◽  
◽  
...  
Keyword(s):  
Stem Cells ◽  
Adipose Tissue ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Somatic Cells ◽  
Embryonic Stem ◽  
Differentiation Potential ◽  
Cytoplasmic Extract ◽  
The Impact ◽  
Dopaminergic Differentiation

The extract of pluripotent stem cells induces dedifferentiation of somatic cells with restricted plasticity. In this study, we used the extract of human embryonic stem cells (hESC) to dedifferentiate adipose tissue-derived stem cells (ADSCs) and examined the impact of this reprogramming event on dopaminergic differentiation of the cells. For this purpose, cytoplasmic extract of ESCs was prepared by repeated cycles of freezing and thawing. The plasma membrane of hADSCs was reversibly permeabilized by Streptolysin O (SLO), exposed to hESC extract and resealed by CaCl2-containing medium. As revealed by qPCR analysis, expression of OCT4, SOX2, NANOG, LIN28A and KLF4 mRNAs were downregulated in the ADSCs one week after extract incubation, while all except for KLF4 were upregulated at the end of second week. For dopaminergic differentiation, control and reprogrammed ADSCs were induced by serum-free neurobasal medium containing B27 and a cocktail of SHH, FGF8, bFGF and BDNF for 12 days. After differentiation, expression levels of some neuronal and dopaminergic-related genes, including PAX6, NESTIN, NEFL, GLI1, LMXB1, EN1, NURR1 and TH, were significantly increase in the reprogrammed ADSCs compared to the control group. As a whole, two weeks after reprogramming by ESC extract, ADSCs showed an improved dopaminergic differentiation potential. These findings suggest that the cytoplasmic extract of hESCs is containing some regulatory factors which induce the expression of pluripotency-associated markers in somatic cells and that the exposure to ESC extract may serve as a simple and rapid strategy to enhance plasticity of somaticstem cells for cell replacement therapy purposes.

Derivation of SSEA4-/CD73+ Mesenchymal Stem Cells from Human Embryonic Stem Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2579-2579
Author(s):  
Parul Trivedi ◽  
Peiman Hematti
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Clinical Applications ◽  
Differentiation Potential ◽  
Hla Class I Antigens ◽  
Undifferentiated Hescs

Abstract Human embryonic stem cells (hESCs) could potentially provide a renewable source of different types of cells for cell therapy applications. Recently, mesenchymal stem cells (MSCs) have been derived from hESCs either through co-culturing with murine OP9 bone marrow stromal cell line or directly from hESCs without co-culturing with OP9 cells. Although the latter methodology is clinically advantageous over co-culturing with an animal cell layer those mesenchymal cells were reported to be positive for SSEA4. SSEA4 is a marker of undifferentiated hESCs and thus the presence of this marker on hESC-derived cells could potentially be problematic for clinical applications. We have recently achieved a novel and reproducible methodology for deriving a pure population of SSEA4-/CD73+ MSCs from federally approved hESC lines H1 and H9. To initiate the differentiation of hESCs to MSCs, we cultured undifferentiated hESCs on matrigel plates in murine embryonic fibroblast conditioned media with media changes every 3 days. Under these culture conditions a portion of embryonic stem cells differentiated into fibroblast looking cells. Through a multi-step process which involved the use of a culture methodology similar to what is being used to culture bone marrow (BM)-derived MSCs, and passaging cultured cells at defined time points we were able to derive a pure population of cells that were uniformly positive for MSC marker CD73 in about a 4-weeks period. These cells had fibroblast/mesenchymal looking morphology, and expressed cell surface marker antigens similar to what has been reported for adult human BM-derived MSCs: they are positive for CD29, CD44, CD54, CD71, CD90, glycophorin A, CD105, and were negative for hematopoietic markers such as CD34 and CD45. Similar to adult BM-derived MSCs these cells express HLA class-I antigens but not class-II antigens. Using established differentiation protocols we could differentiate the hESC-derived CD73+ MSCs into adipocytes, osteocytes, and chondrocytes as verified by immunohistochemistry and RT-PCR assays. So far we have grown these CD73+ MSCs up to passages 15–18. These cells retained their differentiation potential, and were karotypically normal when tested at passage 12. Most importantly, we did not observe any MSCs that were double positive for CD73 and SSEA4 antigen at any time point during our experiments. MSCs from a variety of fetal and adult sources are in various stages of clinical trials with some encouraging preliminary results. Our hESC-derived MSCs that are very similar to adult BM-derived MSCs regarding their growth and morphologic properties, immunophenotypic characteristics, differentiation potential, and importantly are devoid of hESC marker SSEA4 could potentially provide a novel source of MSCs for clinical applications.

Stage-Specific Role for Shh in Dopaminergic Differentiation of Human Embryonic Stem Cells Induced by Stromal Cells

2010 ◽  
Vol 19 (1) ◽  
pp. 71-82 ◽  
Author(s):  
Anna Maria Swistowska ◽  
Alexandre Bettencourt da Cruz ◽  
Yi Han ◽  
Andrzej Swistowski ◽  
Ying Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Stromal Cells ◽  
Embryonic Stem ◽  
Specific Role ◽  
Dopaminergic Differentiation

Human Embryonic Stem Cells May Display Higher Resistance to Genotoxic Stress as Compared to Primary Explanted Somatic Cells

2008 ◽  
Vol 17 (3) ◽  
pp. 599-608 ◽  
Author(s):  
Kumar Jayaseelan Vinoth ◽  
Boon Chin Heng ◽  
Anuradha Poonepalli ◽  
Birendranath Banerjee ◽  
Lakshmidevi Balakrishnan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Somatic Cells ◽  
Embryonic Stem ◽  
Genotoxic Stress

scholarly journals Assessing the impact of minimizing arginine conversion in fully defined SILAC culture medium in human embryonic stem cells

PROTEOMICS ◽  
2016 ◽  
Vol 16 (20) ◽  
pp. 2605-2614 ◽  
Author(s):  
Ellen Scheerlinck ◽  
Katleen Steendam ◽  
Simon Daled ◽  
Elisabeth Govaert ◽  
Liesbeth Vossaert ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Culture Medium ◽  
Embryonic Stem ◽  
The Impact

scholarly journals A role for NANOG in G1 to S transition in human embryonic stem cells through direct binding of CDK6 and CDC25A

2009 ◽  
Vol 184 (1) ◽  
pp. 67-82 ◽  
Author(s):  
Xin Zhang ◽  
Irina Neganova ◽  
Stefan Przyborski ◽  
Chunbo Yang ◽  
Michael Cooke ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
S Phase ◽  
Down Regulation ◽  
Phase Regulation ◽  
The Impact ◽  
Phase Entry

In this study, we show that NANOG, a master transcription factor, regulates S-phase entry in human embryonic stem cells (hESCs) via transcriptional regulation of cell cycle regulatory components. Chromatin immunoprecipitation combined with reporter-based transfection assays show that the C-terminal region of NANOG binds to the regulatory regions of CDK6 and CDC25A genes under normal physiological conditions. Decreased CDK6 and CDC25A expression in hESCs suggest that both CDK6 and CDC25A are involved in S-phase regulation. The effects of NANOG overexpression on S-phase regulation are mitigated by the down-regulation of CDK6 or CDC25A alone. Overexpression of CDK6 or CDC25A alone can rescue the impact of NANOG down-regulation on S-phase entry, suggesting that CDK6 and CDC25A are downstream cell cycle effectors of NANOG during the G1 to S transition.

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