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  1. AU="Thomson, Jason J"
  2. AU="Mitra, Kalyan"
  3. AU="Sanjay Desai"
  4. AU=Cox David J AU=Cox David J
  5. AU="Grebenok, Robert J."
  6. AU="Blackburne, Brittney"
  7. AU="Bortoleti, Bruna Taciane da Silva"
  8. AU="Ehrbar, Martin"
  9. AU="Lepre, Davide"
  10. AU="Olszewska, Zuzanna"
  11. AU="Vojta, Leslie"
  12. AU=Wickstrom Eric AU=Wickstrom Eric
  13. AU="Gangavarapu, Sridevi"
  14. AU="Hussein, Hazem Abdelwaheb"
  15. AU=Cai Yixin AU=Cai Yixin
  16. AU="Hüls, Anke"
  17. AU="Poondru, Srinivasu"
  18. AU="Coca, Daniel"
  19. AU="Lebeau, Paul"
  20. AU="Dehghani, Sedigheh"
  21. AU="Ishibashi, Kenji"
  22. AU="Xu, Yanhua"
  23. AU="Matera, Katarzyna"
  24. AU="Ait-Ouarab, Slimane"
  25. AU="Nicola, Coppede"
  26. AU="Dewitt, John M"
  27. AU="Sorin M. Dudea"
  28. AU="Tanusha D. Ramdin"
  29. AU="Hao, Zehui"
  30. AU="Chauhan, Aman"

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  1. Artikel: A static self-directed method for generating brain organoids from human embryonic stem cells

    Boisvert, Erin M / Means, Robert E / Michaud, Michael / Thomson, Jason J / Madri, Joseph A / Katz, Samuel G

    Journal of visualized experiments. 2020 Mar. 04, , no. 157

    2020  

    Abstract: Human brain organoids differentiated from embryonic stem cells offer the unique opportunity to study complicated interactions of multiple cell types in a three-dimensional system. Here we present a relatively straightforward and inexpensive method that ... ...

    Abstract Human brain organoids differentiated from embryonic stem cells offer the unique opportunity to study complicated interactions of multiple cell types in a three-dimensional system. Here we present a relatively straightforward and inexpensive method that yields brain organoids. In this protocol human pluripotent stem cells are broken into small clusters instead of single cells and grown in basic media without a heterologous basement membrane matrix or exogenous growth factors, allowing the intrinsic developmental cues to shape the organoid's growth. This simple system produces a diversity of brain cell types including glial and microglial cells, stem cells, and neurons of the forebrain, midbrain, and hindbrain. Organoids generated from this protocol also display hallmarks of appropriate temporal and spatial organization demonstrated by brightfield images, histology, immunofluorescence and real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Because these organoids contain cell types from various parts of the brain, they can be utilized for studying a multitude of diseases. For example, in a recent paper we demonstrated the use of organoids generated from this protocol for studying the effects of hypoxia on the human brain. This approach can be used to investigate an array of otherwise difficult to study conditions such as neurodevelopmental handicaps, genetic disorders, and neurologic diseases.
    Schlagwörter basement membrane ; brain ; embryonic stem cells ; fluorescent antibody technique ; genetic disorders ; growth factors ; histology ; humans ; hypoxia ; neuroglia ; neurons ; organoids ; quantitative polymerase chain reaction ; reverse transcriptase polymerase chain reaction
    Sprache Englisch
    Erscheinungsverlauf 2020-0304
    Umfang p. e60379.
    Erscheinungsort Journal of Visualized Experiments
    Dokumenttyp Artikel
    ZDB-ID 2259946-0
    ISSN 1940-087X
    ISSN 1940-087X
    DOI 10.3791/60379
    Datenquelle NAL Katalog (AGRICOLA)

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  2. Artikel ; Online: A Static Self-Directed Method for Generating Brain Organoids from Human Embryonic Stem Cells.

    Boisvert, Erin M / Means, Robert E / Michaud, Michael / Thomson, Jason J / Madri, Joseph A / Katz, Samuel G

    Journal of visualized experiments : JoVE

    2020  , Heft 157

    Abstract: Human brain organoids differentiated from embryonic stem cells offer the unique opportunity to study complicated interactions of multiple cell types in a three-dimensional system. Here we present a relatively straightforward and inexpensive method that ... ...

    Abstract Human brain organoids differentiated from embryonic stem cells offer the unique opportunity to study complicated interactions of multiple cell types in a three-dimensional system. Here we present a relatively straightforward and inexpensive method that yields brain organoids. In this protocol human pluripotent stem cells are broken into small clusters instead of single cells and grown in basic media without a heterologous basement membrane matrix or exogenous growth factors, allowing the intrinsic developmental cues to shape the organoid's growth. This simple system produces a diversity of brain cell types including glial and microglial cells, stem cells, and neurons of the forebrain, midbrain, and hindbrain. Organoids generated from this protocol also display hallmarks of appropriate temporal and spatial organization demonstrated by brightfield images, histology, immunofluorescence and real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Because these organoids contain cell types from various parts of the brain, they can be utilized for studying a multitude of diseases. For example, in a recent paper we demonstrated the use of organoids generated from this protocol for studying the effects of hypoxia on the human brain. This approach can be used to investigate an array of otherwise difficult to study conditions such as neurodevelopmental handicaps, genetic disorders, and neurologic diseases.
    Mesh-Begriff(e) Brain/cytology ; Brain/metabolism ; Cell Differentiation ; Human Embryonic Stem Cells/cytology ; Humans ; Mesencephalon ; Neurons/cytology ; Organoids/metabolism ; Pluripotent Stem Cells/cytology ; Tissue Culture Techniques
    Sprache Englisch
    Erscheinungsdatum 2020-03-04
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Video-Audio Media
    ZDB-ID 2259946-0
    ISSN 1940-087X ; 1940-087X
    ISSN (online) 1940-087X
    ISSN 1940-087X
    DOI 10.3791/60379
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  3. Artikel ; Online: Comparative transcriptome analysis of hESC- and iPSC-derived lentoid bodies.

    Ali, Muhammad / Kabir, Firoz / Thomson, Jason J / Ma, Yinghong / Qiu, Caihong / Delannoy, Michael / Khan, Shahid Y / Riazuddin, S Amer

    Scientific reports

    2019  Band 9, Heft 1, Seite(n) 18552

    Abstract: The ocular lens serves as an excellent system to investigate the intricate details of development and differentiation. Generation of lentoid bodies or lens-like structures using pluripotent stem cells is important for understanding the processes critical ...

    Abstract The ocular lens serves as an excellent system to investigate the intricate details of development and differentiation. Generation of lentoid bodies or lens-like structures using pluripotent stem cells is important for understanding the processes critical for lens morphogenesis and the mechanism of cataractogenesis. We previously reported the generation of peripheral blood mononuclear cell (PBMC)-originated, induced pluripotent stem cells (iPSCs). Here, we report generation of lentoid bodies from human embryonic stem cells (hESCs) and (PBMC)-originated, iPSCs employing the "fried egg" method with brief modifications. The ultrastructure analysis of hESC- and iPSC-derived lentoid bodies identified closely packed lens epithelial- and differentiating fiber-like cells. In addition, we performed RNA sequencing (RNA-Seq) based transcriptome profiling of hESC- and iPSC-derived lentoid bodies at differentiation day 25. Next-generation RNA sequencing (RNA-Seq) of hESC- and iPSC-derived lentoid bodies detected expression (≥0.659 RPKM) of 13,975 and 14,003 genes, respectively. Comparative transcriptome analysis of hESC- and iPSC-derived lentoid bodies revealed 13,563 (>96%) genes common in both datasets. Among the genes common in both transcriptome datasets, 12,856 (~95%) exhibited a quantitatively similar expression profile. Next, we compared the mouse lens epithelial and fiber cell transcriptomes with hESC- and iPSC-derived lentoid bodies transcriptomes and identified > 96% overlap with lentoid body transcriptomes. In conclusion, we report first-time comparative transcriptome analysis of hESC- and iPSC-derived lentoid bodies at differentiation day 25.
    Mesh-Begriff(e) Aged ; Cell Differentiation/genetics ; Cell Line ; Cellular Reprogramming/physiology ; Gene Expression Regulation, Developmental ; Human Embryonic Stem Cells/physiology ; Humans ; Induced Pluripotent Stem Cells/physiology ; Lens, Crystalline/cytology ; Lens, Crystalline/growth & development ; Leukocytes, Mononuclear/physiology ; Male ; Primary Cell Culture ; RNA-Seq ; Transcriptome/physiology
    Sprache Englisch
    Erscheinungsdatum 2019-12-06
    Erscheinungsland England
    Dokumenttyp Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-019-54258-z
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  4. Artikel ; Online: Dimethadione embryotoxicity in the rat is neither correlated with maternal systemic drug concentrations nor embryonic tissue levels.

    Ozolinš, Terence R S / Weston, Andrea D / Perretta, Anthony / Thomson, Jason J / Brown, Nigel A

    Toxicology and applied pharmacology

    2015  Band 289, Heft 1, Seite(n) 89–97

    Abstract: Pregnant rats treated with dimethadione (DMO), the N-demethylated metabolite of the anticonvulsant trimethadione, produce offspring having a 74% incidence of congenital heart defects (CHD); however, the incidence of CHD has high inter-litter variability ( ...

    Abstract Pregnant rats treated with dimethadione (DMO), the N-demethylated metabolite of the anticonvulsant trimethadione, produce offspring having a 74% incidence of congenital heart defects (CHD); however, the incidence of CHD has high inter-litter variability (40-100%) that presents a challenge when studying the initiating events prior to the presentation of an abnormal phenotype. We hypothesized that the variability in CHD incidence was the result of differences in maternal systemic concentrations or embryonic tissue concentrations of DMO. To test this hypothesis, dams were administered 300 mg/kg DMO every 12h from the evening of gestational day (GD) 8 until the morning of GD 11 (six total doses). Maternal serum levels of DMO were assessed on GD 11, 12, 13, 14, 15, 18 and 21. Embryonic tissue concentrations of DMO were assessed on GD 11, 12, 13 and 14. In a separate cohort of GD 12 embryos, DMO concentrations and parameters of growth and development were assessed to determine if tissue levels of DMO were correlated with these endpoints. Embryos were exposed directly to different concentrations of DMO with whole embryo culture (WEC) and their growth and development assessed. Key findings were that neither maternal systemic concentrations nor tissue concentrations of DMO identified embryos that were sensitive or resistant to DMO in vivo. Direct exposure of embryos to DMO via WEC also failed to show correlations between embryonic concentrations of DMO with developmental outcomes in vitro. We conclude that neither maternal serum nor embryonic tissue concentrations of DMO predict embryonic outcome.
    Mesh-Begriff(e) Animals ; Anticonvulsants/toxicity ; Dimethadione/blood ; Dimethadione/toxicity ; Dose-Response Relationship, Drug ; Embryo Culture Techniques ; Embryo, Mammalian/drug effects ; Embryonic Development/drug effects ; Female ; Gestational Age ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Trimethadione/toxicity
    Chemische Substanzen Anticonvulsants ; Dimethadione (ALU9NPM703) ; Trimethadione (R7GV3H6FQ4)
    Sprache Englisch
    Erscheinungsdatum 2015-11-15
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 204477-8
    ISSN 1096-0333 ; 0041-008X
    ISSN (online) 1096-0333
    ISSN 0041-008X
    DOI 10.1016/j.taap.2015.09.005
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  5. Artikel ; Online: Generation and Proteome Profiling of PBMC-Originated, iPSC-Derived Corneal Endothelial Cells.

    Ali, Muhammad / Khan, Shahid Y / Vasanth, Shivakumar / Ahmed, Mariya R / Chen, Ruiqiang / Na, Chan Hyun / Thomson, Jason J / Qiu, Caihong / Gottsch, John D / Riazuddin, S Amer

    Investigative ophthalmology & visual science

    2018  Band 59, Heft 6, Seite(n) 2437–2444

    Abstract: Purpose: Corneal endothelial cells (CECs) are critical in maintaining clarity of the cornea. This study was initiated to develop peripheral blood mononuclear cell (PBMC)-originated, induced pluripotent stem cell (iPSC)-derived CECs.: Methods: We ... ...

    Abstract Purpose: Corneal endothelial cells (CECs) are critical in maintaining clarity of the cornea. This study was initiated to develop peripheral blood mononuclear cell (PBMC)-originated, induced pluripotent stem cell (iPSC)-derived CECs.
    Methods: We isolated PBMCs and programmed the mononuclear cells to generate iPSCs, which were differentiated to CECs through the neural crest cells (NCCs). The morphology of differentiating iPSCs was examined at regular intervals by phase contrast microscopy. In parallel, the expression of pluripotent and corneal endothelium (CE)-associated markers was investigated by quantitative real-time PCR (qRT-PCR). The molecular architecture of the iPSC-derived CECs and human corneal endothelium (hCE) was examined by mass spectrometry-based proteome sequencing.
    Results: The PBMC-originated, iPSC-derived CECs were tightly adherent, exhibiting a hexagonal-like shape, one of the cardinal characteristics of CECs. The CE-associated markers expressed at significantly higher levels in iPSC-derived CECs at days 13, 20, and 30 compared with their respective levels in iPSCs. It is of importance that only residual expression levels of pluripotency markers were detected in iPSC-derived CECs. Cryopreservation of iPSC-derived CECs did not affect the tight adherence of CECs and their hexagonal-like shape while expressing high levels of CE-associated markers. Mass spectrometry-based proteome sequencing identified 10,575 proteins in the iPSC-derived CEC proteome. In parallel, we completed proteome profiling of the hCE identifying 6345 proteins. Of these, 5763 proteins were identified in the iPSC-derived CECs, suggesting that 90.82% of the hCE proteome overlaps with the iPSC-derived CEC proteome.
    Conclusions: We have successfully developed a personalized approach to generate CECs that closely mimic the molecular architecture of the hCE. To the best of our knowledge, this is the first report describing the development of PBMC-originated, iPSC-derived CECs.
    Mesh-Begriff(e) Aged ; Cell Differentiation/physiology ; Cells, Cultured ; Cryopreservation ; Embryonic Stem Cells/cytology ; Endothelium, Corneal/cytology ; Endothelium, Corneal/metabolism ; Flow Cytometry ; Gene Expression Profiling/methods ; Genetic Markers/genetics ; Humans ; Immunohistochemistry ; Induced Pluripotent Stem Cells/cytology ; Induced Pluripotent Stem Cells/metabolism ; Leukocytes, Mononuclear/cytology ; Leukocytes, Mononuclear/metabolism ; Male ; Mass Spectrometry ; Microscopy, Phase-Contrast ; Middle Aged ; Neural Crest/cytology ; Proteome/genetics ; Real-Time Polymerase Chain Reaction
    Chemische Substanzen Genetic Markers ; Proteome
    Sprache Englisch
    Erscheinungsdatum 2018-06-12
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 391794-0
    ISSN 1552-5783 ; 0146-0404
    ISSN (online) 1552-5783
    ISSN 0146-0404
    DOI 10.1167/iovs.17-22927
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  6. Artikel ; Online: Noninvasive high-resolution ultrasound reveals structural and functional deficits in dimethadione-exposed fetal rat hearts in utero.

    Purssell, Elizabeth / Weston, Andrea D / Thomson, Jason J / Swanson, Terri A / Brown, Nigel A / Ozolinš, Terence R S

    Birth defects research. Part B, Developmental and reproductive toxicology

    2012  Band 95, Heft 1, Seite(n) 35–46

    Abstract: Background: We previously showed dimethadione (DMO), the N-demethylated metabolite of the anticonvulsant trimethadione, induces ventricular septation defects (VSD) and other heart anomalies in rat (Weston et al., 2011). Because of the relationship ... ...

    Abstract Background: We previously showed dimethadione (DMO), the N-demethylated metabolite of the anticonvulsant trimethadione, induces ventricular septation defects (VSD) and other heart anomalies in rat (Weston et al., 2011). Because of the relationship between cardiac structure and function, we hypothesized that DMO-induced structural defects of the heart are associated with in utero functional deficits. To test the hypothesis, the goals were (1) define the parameters for ultrasound in the rat conceptus, and; (2) use ultrasound to identify structural and functional deficits following DMO treatment.
    Methods: Different ultrasound modes (B-mode, M-mode, and Pulse-wave Doppler) using four high-resolution ultrasound transducer heads of varying frequency (25-40 MHz) were tested on gestational day (GD) 14, 15, 16, 17, and 21. Having identified the optimal conditions, pregnant Sprague-Dawley rats were administered six 300 mg/kg doses of DMO every 12 hr beginning at 19:00 hr on GD 8 to generate conceptuses with a high incidence of VSD.
    Results: The three ultrasound modalities were used to identify VSD and several novel and rare structural heart anomalies (cardiac effusions and bifurcated septum) in live rat fetuses. DMO-treated hearts had an array of functional deficits including a decrease in mean heart rate, ejection fraction, and cardiac output and increased incidence of bradycardia and dysrhythmia.
    Conclusions: The ultrasound biomicroscope is an effective tool for the real-time characterization of the structure and function of embryo/fetal rat hearts. DMO causes significant deficits to in utero heart function for up to ten days (GD 21) following its final administration, suggesting long-term or possible permanent changes cardiac function.
    Mesh-Begriff(e) Animals ; Dimethadione/adverse effects ; Female ; Fetus/drug effects ; Fetus/physiopathology ; Heart/drug effects ; Heart/embryology ; Heart/physiopathology ; Heart Function Tests ; Heart Rate/drug effects ; Heart Septal Defects, Ventricular/diagnostic imaging ; Isoflurane/adverse effects ; Myocardial Contraction/drug effects ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Ultrasonics ; Ultrasonography
    Chemische Substanzen Dimethadione (ALU9NPM703) ; Isoflurane (CYS9AKD70P)
    Sprache Englisch
    Erscheinungsdatum 2012-02
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2104792-3
    ISSN 1542-9741 ; 1542-0752 ; 1542-9733 ; 1542-975X
    ISSN (online) 1542-9741
    ISSN 1542-0752 ; 1542-9733 ; 1542-975X
    DOI 10.1002/bdrb.20339
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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    Zs.A 749: Hefte anzeigen Standort:
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  7. Artikel ; Online: Assessment of the Embryonic Stem Cell Test and application and use in the pharmaceutical industry.

    Paquette, Jennifer A / Kumpf, Steven W / Streck, Randal D / Thomson, Jason J / Chapin, Robert E / Stedman, Donald B

    Birth defects research. Part B, Developmental and reproductive toxicology

    2008  Band 83, Heft 2, Seite(n) 104–111

    Abstract: Background: The European Centre for the Validation of Alternative Methods (ECVAM) designed the Embryonic Stem Cell Test (EST) as a tool for classifying developmentally toxic compounds. An in vitro tool to assess developmental toxicity would be of great ... ...

    Abstract Background: The European Centre for the Validation of Alternative Methods (ECVAM) designed the Embryonic Stem Cell Test (EST) as a tool for classifying developmentally toxic compounds. An in vitro tool to assess developmental toxicity would be of great value to the pharmaceutical industry to help with toxicity-associated attrition.
    Methods: ECVAM's EST protocol was used, but employing a different mouse embryonic stem cell (ESC) line and an alternative differentiation medium. A subset of the compounds used to validate the EST assay along with a number of in-house pharmaceutical compounds plus marketed pharmaceutical compounds were used to assess the EST performance with receptor-mediated compounds.
    Results: Our results with ECVAM compounds mirrored ECVAM's. Compounds that were developmentally toxic in vivo were classified by the EST as moderate risk. Overall, the accuracy was 75% with the current set of data and the predictivity of low-, moderate-, and high-risk compounds was 90, 71, and 60% while the precision was 59, 86, and 100%, respectively. Interestingly, a number of the non-developmentally toxic compounds had values for the 3T3 IC(50) values, which were lower than the ESC IC(50) and ID(50), a situation not taken into account by ECVAM when designing the EST algorithm.
    Conclusions: The assay as currently constructed has a significant false-positive rate (approximately 40%), but a very low false-negative rate (approximately 7%). Additional moderate- and high-risk compounds need to be assessed to increase confidence, accuracy, and understanding in the EST's predictivity.
    Mesh-Begriff(e) Animal Testing Alternatives/methods ; Animals ; BALB 3T3 Cells ; Cell Differentiation/drug effects ; Cell Survival/drug effects ; Cells, Cultured ; Drug Industry/methods ; Embryonic Stem Cells/drug effects ; Embryonic Stem Cells/physiology ; Mice ; Mice, Inbred DBA ; Toxicity Tests/methods
    Sprache Englisch
    Erscheinungsdatum 2008-04
    Erscheinungsland United States
    Dokumenttyp Evaluation Studies ; Journal Article
    ZDB-ID 2104792-3
    ISSN 1542-9741 ; 1542-0752 ; 1542-9733 ; 1542-975X
    ISSN (online) 1542-9741
    ISSN 1542-0752 ; 1542-9733 ; 1542-975X
    DOI 10.1002/bdrb.20148
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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