scholarly journals In Vivo Human PSC-Derived Intestinal Organoids to Study Stem Cell Maintenance

2020 ◽  
pp. 201-214
Author(s):  
Simon Vales ◽  
Holly M. Poling ◽  
Nambirajan Sundaram ◽  
Michael A. Helmrath ◽  
Maxime M. Mahe
Keyword(s):  
Stem Cell ◽  
Stem Cell Maintenance ◽  
Intestinal Organoids ◽  
Cell Maintenance

scholarly journals New in vivo microRNA biotechnology reveals specific roles for the miR‐200 family in dental stem cell maintenance

2019 ◽  
Vol 33 (S1) ◽  
Author(s):  
Mason E Sweat ◽  
Yan Yan Sweat ◽  
Steven Eliason ◽  
Wenjie Yu ◽  
Huojun Cao ◽  
...  
Keyword(s):  
Stem Cell ◽  
Stem Cell Maintenance ◽  
Dental Stem Cell ◽  
Cell Maintenance

scholarly journals The Transcription Regulator Patz1 Is Essential for Neural Stem Cell Maintenance and Proliferation

2021 ◽  
Vol 9 ◽  
Author(s):  
Sara Mancinelli ◽  
Michela Vitiello ◽  
Maria Donnini ◽  
Francesca Mantile ◽  
Giuseppe Palma ◽  
...  
Keyword(s):  
Stem Cell ◽  
Brain Development ◽  
Ex Vivo ◽  
Embryonic Stem ◽  
Adult Brain ◽  
Transcription Regulator ◽  
Stem Cell Maintenance ◽  
Stem And Progenitor Cells ◽  
Cell Maintenance

Proper regulation of neurogenesis, the process by which new neurons are generated from neural stem and progenitor cells (NS/PCs), is essential for embryonic brain development and adult brain function. The transcription regulator Patz1 is ubiquitously expressed in early mouse embryos and has a key role in embryonic stem cell maintenance. At later stages, the detection of Patz1 expression mainly in the developing brain suggests a specific involvement of Patz1 in neurogenesis. To address this point, we first got insights in Patz1 expression profile in different brain territories at both embryonic and postnatal stages, evidencing a general decreasing trend with respect to time. Then, we performed in vivo and ex vivo analysis of Patz1-knockout mice, focusing on the ventricular and subventricular zone, where we confirmed Patz1 enrichment through the analysis of public RNA-seq datasets. Both embryos and adults showed a significant reduction in the number of Patz1-null NS/PCs, as well as of their self-renewal capability, compared to controls. Consistently, molecular analysis revealed the downregulation of stemness markers in NS/PCs derived from Patz1-null mice. Overall, these data demonstrate the requirement of Patz1 for NS/PC maintenance and proliferation, suggesting new roles for this key transcription factor specifically in brain development and plasticity, with possible implications for neurodegenerative disorders and glial brain tumors.

scholarly journals Elongator is required for root stem cell maintenance by regulating SHORT ROOT transcription

2018 ◽  
Author(s):  
Linlin Qi ◽  
Xiaoyue Zhang ◽  
Huawei Zhai ◽  
Jian Liu ◽  
Fangming Wu ◽  
...  
Keyword(s):  
Stem Cell ◽  
Rna Polymerase Ii ◽  
Transcription Elongation ◽  
Direct Interaction ◽  
Short Root ◽  
Stem Cell Maintenance ◽  
Radial Patterning ◽  
Root Stem Cell ◽  
Cell Maintenance

AbstractSHORTROOT (SHR) is essential for stem cell maintenance and radial patterning in Arabidopsis thaliana roots, but how its expression is regulated is still unknown. Here, we report that Elongator regulates the transcription of SHR. The depletion of Elongator drastically reduced SHR expression and led to defective root stem cell maintenance and radial patterning. The importance of the nuclear localization of Elongator for its functioning, together with the insensitivity of the elp1 mutant to the transcription elongation inhibitor 6-azauracil and the direct interaction of the ELP4 subunit with the C-terminal domain of RNA polymerase II (RNAPII CTD), support the notion that Elongator plays important roles in transcription elongation. Indeed, we found that ELP3 associates with the pre-mRNA of SHR and that mutation of Elongator reduces the enrichment of RNAPII on the SHR gene body. Moreover, Elongator interacted in vivo with SUPPRESSOR OF Ty4 (SPT4), a well-established transcription elongation factor that was recruited to the SHR locus. Together, these results demonstrate that Elongator acts in concert with SPT4 to regulate the transcription of SHR.

Lentiviral Transduction of Patient Derived Leukaemic Blasts Allowing In Vivo Bioluminescent Monitoring in An NSG Model of Leukaemia Stem Cell Maintenance,

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 4000-4000 ◽  
Author(s):  
Simon Bomken ◽  
Lars Buechler ◽  
Klaus Rehe ◽  
Frida Ponthan ◽  
Helen Blair ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Stem Cell ◽  
Fluorescent Protein ◽  
Imaging System ◽  
Conflicts Of Interest ◽  
Stem Cell Maintenance ◽  
Serial Transplantation ◽  
Cell Maintenance

Abstract Abstract 4000 Serial transplantation of patient derived acute lymphoblastic leukaemia (ALL) blasts continues to contribute to our understanding of the biology of leukaemia stem cells (LSC). Refinement of techniques, and in particular intrafemoral injection and development of the highly immunocompromised NOD/scid IL2Rγ null (NSG) mouse have demonstrated B precursor ALL propagating cells to be both common and present in diverse immunophenotypes. These studies must now be complemented by interrogation of the biological pathways underpinning leukaemia stem cell behaviour and clonal propagation in ALL. We have developed a lentiviral based approach to such studies, using the transfer vector pSLIEW, encoding both enhanced green fluorescent protein (EGFP) and firefly luciferase (luc). We have recently replaced a bone marrow stromal feeder based transduction protocol with a feeder free protocol, removing the risk of co-transduction of feeder cells. Using the feeder free protocol, we have achieved transduction of primary (n=4) and primograft (n=3) material with between 13.0 and 51.4% eGFP positive cells. Transplantation of transduced cells by intrafemoral injection into NSG mice resulted in engraftment and disease dissemination. This process was monitored using an IVIS Spectrum bioluminescence imaging system. This technique demonstrated progression of disease to the contralateral femur, spleen, CNS and vertebrae. Disease progression was also monitored by serial bone marrow punctures and 5-colour flow cytometry, which demonstrated no immunophenotypic bias amongst the transduced cells. Flow cytometry of harvested bone marrow and spleen showed between 5.5% and 10.2% eGFP positive cells representing only a moderate decrease from 26.3% eGFP positivity at initial transplantation. This confirms the relative resistance of the SFFV promoter to silencing, making this approach suitable for serial transplantation. Harvested bone marrow and splenic cells were re-transplanted at 5.5 × 103 – 1.0 × 104 SLIEW+ cells per mouse (total 1 × 105 cells transplanted). Bioluminescent imaging has shown engraftment and dissemination of leukaemia within five weeks, confirming transduction of the leukaemia stem cell compartment. Further development of the pSLIEW vector to include shRNA sequences now offers the potential for functional studies using patient derived material, transduced with a single lentivector construct and serially engrafted in the NSG assay for leukaemic stem cell maintenance. We believe that this approach will allow us to investigate the genetic programmes underpinning leukaemia stem cell self-renewal. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Non-canonical function of FIP200 is required for neural stem cell maintenance and differentiation by limiting TBK1 activation and p62 aggregate formation

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Hang Liu ◽  
Chenran Wang ◽  
Fei Yi ◽  
Syn Yeo ◽  
Michael Haas ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Neural Stem Cell ◽  
Kinase Activity ◽  
Aggregate Formation ◽  
Canonical Function ◽  
Stem Cell Maintenance ◽  
Autophagy Gene ◽  
Cell Maintenance

AbstractFIP200 is an essential autophagy gene implicated in the regulation of postnatal neural progenitor/stem cells (NSCs). However, the contribution of FIP200’s canonical-autophagy function and its non-canonical functions to postnatal NSC maintenance remains unclear. Utilizing a recently generated Fip200-4A allele that specifically impairs FIP200’s canonical-autophagy function, we found that non-canonical functions of FIP200 was required for regulation of mouse NSC maintenance and neurogenesis in vivo. Ablating the non-canonical functions of FIP200, but not its autophagy function, increased TBK1 activation and p62 phosphorylation at S403 in NSCs. Phosphorylation of p62 was dependent on TBK1 kinase activity and increased the propensity of p62 aggregate formation specifically in FIP200-null NSCs. Accordingly, inhibition of TBK1 by amlexanox reduced p62 aggregates and restored NSC maintenance and differentiation in Fip200hGFAP cKO mice. These results reveal a mechanism for the non-canonical functions of FIP200 in NSC maintenance and differentiation by limiting TBK1 activation and subsequently, p62 aggregate formation.

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