scholarly journals Retroviral Gene Expression in Spermatogonial Stem Cells during Long-term Culture

2007 ◽  
Vol 20 (7) ◽  
pp. 1015-1022
Author(s):  
D. K. Jeong ◽  
Michael D. Griswold
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Spermatogonial Stem Cells ◽  
Long Term Culture

Long-Term Culture and Transplantation of Spermatogonial Stem Cells from BALB/c Mice

2010 ◽  
Vol 191 (5) ◽  
pp. 372-381 ◽  
Author(s):  
Fujin Shen ◽  
Ci Zhang ◽  
Hongyun Zheng ◽  
Yunhe Xiong ◽  
Xi Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Spermatogonial Stem Cells ◽  
Long Term Culture

Activity of OTX015 (MK-8628), a BET-Bromodomain Inhibitor, in Acute Myeloid Leukemia (AML) Progenitor Cells

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 2588-2588 ◽  
Author(s):  
Louise Roulin ◽  
Ashfaq Ali ◽  
Aline Masse ◽  
Marie-Magdelaine Coudé ◽  
Dominique Bluteau ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Progenitor Cells ◽  
Cell Lines ◽  
Research Funding ◽  
Low Dose ◽  
Prolonged Exposure ◽  
Target Concentration ◽  
Long Term Culture

Abstract CONTEXT: Eradication of leukemic progenitor cells, defined by functional assays such as long-term culture (leukemic long-term culture initiating cells [L-LTC-IC]) is the goal of therapy in AML. Bromodomain and ExtraTerminal (BET) proteins are epigenetic readers that regulate the expression of genes with super-enhancers, including CMYC. BET inhibitors (BETi) such as JQ1 induce proliferation arrest and apoptosis in murine models of AML, in human AML cell lines and primary blasts. Their activity in human leukemic progenitors has not yet been reported. OTX015 (MK-8626) is an orally available BETi that can be safely administered to patients with a continuous low-dose regimen (Dombret et al. Blood. 2014). Single-dose exposure to OTX015 induces gene expression modulation characteristic of bromodomain inhibition, including downregulation of CMYC and upregulation of HEXIM1, inhibiting the viability of AML cell lines, and inducing apoptosis in primary AML blasts (Coudé et al. Oncotarget. 2015). To address the activity of OTX015 on leukemic progenitors, we analyzed (A) the clonogenicity of AML cell lines and (B) the frequency of primary L-LTC-IC after repeated low-dose exposure to OTX015. METHODS: (A) Five AML cell lines (OTX015 IC50 60 - 10,000 nM) were studied: OCI-AML3, NOMO-1, HL-60, KG1a and K562. After 24h starvation, OTX015 or vehicle (DMSO) was added daily to the culture medium for 3 days at various concentrations. After 96h, cells were assessed for gene expression by RT-qPCR and seeded in methycellulose. Colonies were scored after 14 days. (B) Bone-marrow mononuclear cells (BMNC) from AML patients obtained at diagnosis after informed consent were cultured for three weeks in a niche-like hypoxic milieu shown to maintain leukemic stem cells (Griessinger et al. Stem Cells Transl Med. 2014). OTX015 200 nM or DMSO was added weekly. This concentration is in the range of trough concentrations achievable at the MTD of OTX015 in phase I trials. Residual leukemic cells were sorted and plated on methylcellulose. Colonies were scored after 14 days. The resulting L-LTC-IC frequency was reported relative to the number of BMNC initially seeded. RESULTS: (A) To dissect the effect of OTX015 on AML progenitors from that on the leukemic bulk, we determined for each cell line a maximal OTX015 concentration that could be administered repeatedly for 3 days without significantly impairing proliferation or viability (MTT) at day 4 of culture (referred as low-dose concentration). As expected, this target concentration, ranging from 50 to 500 nM, was lower in cell lines with low OTX015 IC50. This prolonged low-dose exposure to OTX015 recapitulated BETi-associated gene expression changes including CMYC downregulation and HEXIM1 upregulation in all cell lines, and significantly reduced clonogenicity compared to DMSO in 4/5 cell lines, but not in NPM1-mutated OCI-AML3 cells (IC50: 60 nM, target concentration 50 nM), despite modulation of CMYC and HEXIM1 expression. Overall, there was no correlation between the level of CMYC repression and clonogenicity. Transcriptome analyses are ongoing to identify gene expression changes specifically associated with inhibition of clonogenicity. (B) L-LTC-IC frequency after prolonged exposure to 200 nM OTX015 was determined in specimens from 11 AML patients with variable oncogenetics. L-LTC-IC frequency was reduced in 5/11 patients, reaching statistical significance in 3 cases; OTX015 reduced L-L-LTC-IC in 3 of 4 NPM1-mutated samples, but not in any of the 3 patients with high-risk cytogenetics. No clear correlation was found between induction of apoptosis on primary blasts after short-term, and L-LTC-IC reduction after long-term 200nM OTX015 exposure respectively. Patients' samples number is being extended to identify oncogenetic predictors of L-LTC-IC reduction. CONCLUSION: Our results suggest that in AML cell lines or primary samples, prolonged exposure to low concentrations of the clinically-available BET inhibitor OTX015 results in activity against leukemic progenitors independent of induction of proliferation arrest or apoptosis in blasts. Molecular mechanisms and oncogenic markers of this activity are being investigated. These results warrant clinical investigation of the anti-leukemic properties of prolonged low-dose OTX015 administration. Disclosures Riveiro: Oncoethix: Research Funding; OTD: Employment. Herait:Oncoethix: Other: shareholder; Oncoethix: Other: Chief medical officer; Oncoethix: Other: shareholder. Dombret:Oncoethix: Research Funding. Itzykson:Oncoethix: Research Funding.

scholarly journals Expansion and long-term culture of human spermatogonial stem cells via the activation of SMAD3 and AKT pathways

2015 ◽  
Vol 240 (8) ◽  
pp. 1112-1122 ◽  
Author(s):  
Ying Guo ◽  
Linhong Liu ◽  
Min Sun ◽  
Yanan Hai ◽  
Zheng Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Spermatogonial Stem Cells ◽  
Long Term Culture

Genetic and epigenetic stability of human spermatogonial stem cells during long-term culture

2014 ◽  
Vol 102 (6) ◽  
pp. 1700-1707.e1 ◽  
Author(s):  
Bita Nickkholgh ◽  
S. Canan Mizrak ◽  
Saskia K.M. van Daalen ◽  
Cindy M. Korver ◽  
Hooman Sadri-Ardekani ◽  
...  
Keyword(s):  
Stem Cells ◽  
Spermatogonial Stem Cells ◽  
Long Term Culture ◽  
Genetic And Epigenetic Stability

scholarly journals Culture of spermatogonial stem cells and use of surrogate sires as a breeding technology to propagate superior genetics in livestock production: A systematic review

2021 ◽  
pp. 3235-3248
Author(s):  
Wilkister Nakami ◽  
Ambrose Ng'eno Kipyegon ◽  
James Nguhiu-Mwangi ◽  
Christian Tiambo ◽  
Stephen Kemp
Keyword(s):  
Systematic Review ◽  
Stem Cells ◽  
Data Extraction ◽  
Transgenic Animals ◽  
Spermatogonial Stem Cells ◽  
Published Data ◽  
Long Term Culture ◽  
Future Possibility

Background and Aim: Spermatogonial stem cells (SSCs) have previously been isolated from animals' testes, cultured in vitro, and successfully transplanted into compatible recipients. The SSC unique characteristic has potential for exploitation as a reproductive tool and this can be achieved through SSC intratesticular transplantation to surrogate sires. Here, we aimed at comprehensively analyzing published data on in vitro maintenance of SSC isolated from the testes of livestock animals and their applications. Materials and Methods: The literature search was performed in PubMed, Science Direct, and Google Scholar electronic databases. Data screening was conducted using Rayyan Intelligent Systematic Review software (https://www.rayyan.ai/). Duplicate papers were excluded from the study. Abstracts were read and relevant full papers were reviewed for data extraction. Results: From a total of 4786 full papers screened, data were extracted from 93 relevant papers. Of these, eight papers reported on long-term culture conditions (>1 month) for SSC in different livestock species, 22 papers on short-term cultures (5-15 days), 10 papers on transfection protocols, 18 papers on transplantation using different methods of preparation of livestock recipients, and five papers on donor-derived spermatogenesis. Conclusion: Optimization of SSC long-term culture systems has renewed the possibilities of utilization of these cells in gene-editing technologies to develop transgenic animals. Further, the development of genetically deficient recipients in the endogenous germline layer lends to a future possibility for the utilization of germ cell transplantation in livestock systems.

scholarly journals Cardiac Specific Gene Expression Changes in Long Term Culture of Murine Mesenchymal Stem Cells

2011 ◽  
Vol 4 (2) ◽  
pp. 143-148 ◽  
Author(s):  
Hannah M. Hodgkiss-Geere ◽  
David J. Argyle ◽  
Brendan M. Corcoran ◽  
Bruce Whitelaw ◽  
Elspeth Milne ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Specific Gene ◽  
Long Term Culture ◽  
Specific Gene Expression

Gene expression during long-term culture of mesenchymal stem cells obtained from patients with amyotrophic lateral sclerosis

2012 ◽  
Vol 6 (4) ◽  
pp. 342-353 ◽  
Author(s):  
Mi Ran Choi ◽  
Nando Dulal Das ◽  
Kyoung Hwa Jung ◽  
Seung Hyun Kim ◽  
Hyun Young Kim ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Amyotrophic Lateral Sclerosis ◽  
Mesenchymal Stem Cells ◽  
Long Term Culture ◽  
Lateral Sclerosis
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