Guided differentiation of ES cell into mesenchymal stem cell
|
|
- Sharleen Paul
- 6 years ago
- Views:
Transcription
1 Guided differentiation of ES cell into mesenchymal stem cell Takumi Era Division of Molecular Neurobiology Institute of Molecular Embryology and Genetics, Kumamoto University
2 Differentiation pathways of in vitro ES cell culture Neuroepithelium Sox1 + Sox1 - PDGFRα - PDGFRα + Mesenchymal Stem Cell Adipocyte Osteocyte Chondrocyte ES cell PDGFRα + VEGFR2 + PDGFRα + VEGFR2 + PDGFRα + VEGFR2 - Paraxial Mesoderm 1. To visualize the intermediates Immature New Mesoderm surface markers; PDGFRα and CXCR4 2. To establish the defined conditions PDGFRα -- Lateral Factors; Activin and VEGFR2 BMP4 + Mesoderm 3. To find the new molecules involved Primitive in Endoderm development Adipocyte Osteocyte Chondrocyte Blood cell Endothelial cell CXCR4 + ECD + Definitive Endoderm Hepatocyte Gut cell GSC + ECD + Mesendoderm Nishikawa and Era, Nature Rev. Mol. Cell. Biol., 27 CXCR4 + ECD - Axial Mesoderm Osteocyte
3 Gene expressions in the intermediates Expression Intensity Gene 1 Gene 2 Eomeso Sox17 Brachyury foregut Midgut E8.5 mouse embryo ES cell PDGFRα+ VEGFR2+ Mesoderm PDGFRα+ VEGFR2- PDGFRα- VEGFR2+ Day 1 Day 2 Day 3 Activin added Day 4 Day 5 Gsc +,E-cad + Endoderm differentiation Endoderm cell line ada and Era, Development, 25 akebe and Era, Dev. Biol. 26 ES cell-derived samples
4 Mixl1 expression restore the defect of endoderm by Eomeso KD Endoderm marker, Sox17-hCD Control Eomesodermin Knock-down Vector only Vector only Mixl1-overexpressed Endoderm Mesendoderm marker, Goosecoid-GFP Eomesodermin Mixl1 Sox17 Izumi and Era, Stem Cells, 27
5 Differentiation pathways of in vitro ES cell culture Neuroepithelium Sox1 + Sox1 - PDGFRα - PDGFRα + Mesenchymal Stem Cell Adipocyte Osteocyte Chondrocyte ES cell PDGFRα + VEGFR2 + Immature Mesoderm PDGFRα + VEGFR2 - Paraxial Mesoderm 4. To answer an important question in developmental PDGFRα -- Lateral VEGFR2 biology + Mesoderm Adipocyte Osteocyte Chondrocyte Blood cell Endothelial cell PDGFRα + VEGFR2 + Primitive Endoderm CXCR4 + ECD + Definitive Endoderm Hepatocyte Gut cell GSC + ECD + Mesendoderm Nishikawa and Era, Nature Rev. Mol. Cell. Biol., 27 CXCR4 + ECD - Axial Mesoderm Osteocyte
6 What is Mesenchymal stem cell (MSC)? Definition Fibroblastic morphology Adipocytes Self-renewal Chondrocytes Osteocytes Marshak, D et al, Science, 1999
7 Problems of MSCs Research Utilities Multipotency for differentiation Prevention from GVHD in transplantation Support for regeneration of cardiac muscles Problems Developemtal pathway is unclear Specific surface markers are undefined Transplantation methods are not established ES cell study are useful for understanding what MSC is Aims: To identify MSC progenitors and to define MSC differentiation pathway
8 Two distinct conditions induce adipocytes in in vitro ES cell culture Condition A (-RA) Day Day 5 Day 9 -RA + Insulin, dexamethasone, IBMX and troglitazone Day 18 Adipogenesis ES cells Monolayer Culture Sakurai, Era and Nishikawa et al, Stem Cells, 26 Condition B (+RA) +RA Day On day 2 and 3 Day 5 Day 9 + Insulin, dexamethasone, IBMX and troglitazone Monolayer Culture Dani C. and Austin S. et al, Development, 1997 Oil red O staining
9 Both condition induces the PDGFRα + adipocytic progenitors Condition A (-RA) Day 4 Day 5 Day 9 Adipogenesis 8 4.1% 8 2.4% % TG:26 Condition B (+RA) % % % TG: 327 Mesoderm and Mesenchymal Marker, PDGFRα Oil red O staining
10 Condition B induces PDGFRα + mesenchymal cells Condition A (-RA) Day 4 Day 5 Day % 8 2.4% % Mesodermal cells 4 2 Brachyury Mesoderm Markers Mesp2 Mesogenin OB-CAD Condition B (+RA) Mesenchymal Markers Vimentin PDGFRβ % % Mesenchymal cells % βactin RT-PCR Mesoderm and Mesenchymal Marker, PDGFRα
11 ES cell-derived PDGFRα + Mesenchymal stem cells Markers Cell Growth Adipocytes Oil red O staining PPARγ Adiponectin 19 PDGFRα+ mesenchymal cells Fibroblastic morphology Cell number 14 9 PDGFRα+ mesoderm cells Cloning Osteocytes Alizalin red staining Osteopontin Osteocalcin Passage Condition A-derived Condition B-derived Chondrocytes Alucian blue staining x Col2a1 Sox9
12 What kinds of cells are progenitors for MSCs? +RA Mesenchymal stem cells ES cells?? PDGFRα + adipocytes chondrocytes osteocytes
13 RA treatment induces neuro-epithelial cells Condition A Condition B -RA +RA Mesoderm Day Brachyury Mesogenin Endoderm Foxa2 GATA4 Ectoderm Otx2 Sox1 Neurogenin1 β-actin RT-PCR
14 Visualization of neuroepithelium as Sox1-GFP + cells Neuroepithelium Sox1 Expression 8.5dpc 9.5dpc Neural Tube ISH Pevny et al. Development, 1998 Sox1-GFP Knock-in ES cells Sox1 Promoter GFP J. Aubert et al. PNAS, 23
15 Neuro-epithelium is a origin of ES cell-derived MSC Condition B (+RA) Day 4 6 days after purification Adipogenesis Mesenchyme marker PDGFRα.8.5 Sox1 + Purification 48 PDGFRα Sox1-GFP Neuroepithelium marker Sox1-GFP
16 Differentiation pathway from ES cell to MSC Neuro-epithelial cells Mesenchymal stem cells ES cells +RA Sox1+ PDGFRα- Sox1- PDGFRα+ adipocytes chondrocytes osteocytes
17 4N tetraploid embryo Tetraploid chimera Only ES cells can contribute to embryo. 2N ES cell carrying GFP marker E9.5 x Review by Tam, PP. and Rossant, J. Development, 23 Sox1-GFP Tetraploid chimera Sox1/Nestin/TO-PRO
18 Visualization of Sox1 + cells in mouse embryos Adipogenesis Sox1-GFP tetraploid chimera FACS analysis 41 4 Trunk region PDGFRα Mesoderm Neuroepithelium Osteogenesis 31 Cloning E9.5 Mouse embryo Sox1-GFP Neuro-epithelium is the earliest origin of MSCs Chondrogenesis Takashima, Era and Nishikawa Cell, 27
19 Development of Neural Crest Cells They appear in the border between nuero-epithelium and epidermis and migrates from dorsal to ventral regions. Bone, cartilage and neuron in head, and synpathetic neurons and pigment cells in trunk Neuroepithelium Neural crest cells Gilbert, Developmental Biology
20 Tracing of Sox1 + neuro-epithelial and P + neural crest cells Sox1-Cre Knock-in mouse Cre gene is specifically expressed in Sox1 + neuro-epithelium cells P-Cre transgenic mouse Cre gene is specifically expressed in P + neural crest cells Rosa26-STOP-YFP mouse Rosa-p loxp Cre recombinase Rosa-p STOP YFP loxp YFP YFP marker are constitutively expressed during mouse development
21 YFP expression in Sox1-Cre and P-Cre embryos 21 1 Sox1-Cre euroepithelium) P-Cre Neural crest) FACS Analysis PDGFRα PDGFRα 13 YFP E14.5 embryos YFP
22 Neuroepithelium-and neural crest-derived MSCs in mouse embryo Colonies of MSCs, Colony Forming Unit-Fibroblast (CFU-F) Sox1-Cre (neuroepithelium-derived) P-Cre(Neural crest-derived) + + ox1/yfp PDGFRα P/YFP PDGFRα Neuroepithliumderived Neural crestderived Neuroepithelium- and neural crest-derived PDGFRα + MSCs are present in the trunk of E14.5 mouse embryos
23 E14.5 Sox1 + -derived PDGFRα + cells shows a multipotency and a sustained growth in vitro Cell Morphology:Fibroblastic Gene expression PDGFRα Single cell sorting βactin PDGFRα OBCAD PDGFRβ Vimentin Slug Snail Passage 18 RT-PCR Sox1-Cre, Rosa-YFP Differentiation Potential of clonal cell lines Adipogenesis Osteogenesis Chondrogenesis
24 MSCs activity of neonate Sox1 + -derived PDGFRα + cells FACS pattern of Femur bone (from P to P7) 3 CFU-F assay YFP + YFP - YFP - YFP + PDGFRα + PDGFRα + PDGFRα - PDGFRα - PDGFRα 2 1 Single cell Sorting Sox1-Cre YFP Fibroblastic Morphology.3 Differentiation activity of clonal cell lines Adipogenesis Osteogenesis Chondrogenesis Passage 18
25 NE-derived MSC are reduced after birth Proportion of PDGFRα + population %.15 4-week old Neonates. P<.1.5 YFP + PDGFRα + YFP + PDGFRα -
26 New differentiation pathway of MSCs Neuro-epithelium Neural crest MSCs Sox1 + PDGFRα - P + Sox1 - PDGFRα + Takashima, Era and Nishikawa, Cell, 27
27 Two developmental waves of MSCs Neuro-epithelium supplies the earliest and transient wave of MSCs Neuroepithelium origin Unknown origin Mesoderm? MSC activity Embryo After Birth Development
28 Acknowledgements RIKEN CDB Stem Cell Biology Group Yasuhiro Takashima Shin-Ichi Nishikawa Cambridge University Austin Smith Animal Resources and Genetic Engineering Kazuki Nakao
NPTEL Biotechnology Tissue Engineering. Stem cells
Stem cells S. Swaminathan Director Centre for Nanotechnology & Advanced Biomaterials School of Chemical & Biotechnology SASTRA University Thanjavur 613 401 Tamil Nadu Joint Initiative of IITs and IISc
More informationPRIME-XV Cell Therapy Products by
PRIME-XV Cell Therapy Products by Product List 2017 TRINOVA BIOCHEM now distributes the PRIME-XV Cell Therapy line by Irvine Scientific in Germany, Austria and Switzerland. Irvine Scientific is a worldwide
More informationXeno-Free Systems for hesc & hipsc. Facilitating the shift from Stem Cell Research to Clinical Applications
Xeno-Free Systems for hesc & hipsc Facilitating the shift from Stem Cell Research to Clinical Applications NutriStem Defined, xeno-free (XF), serum-free media (SFM) specially formulated for growth and
More informationPRIME-XV Cell Therapy Products by
PRIME-XV Cell Therapy Products by Product List 2017 TRINOVA BIOCHEM now distributes the PRIME-XV Cell Therapy line by Irvine Scientific in Germany, Austria and Switzerland. Irvine Scientific is a worldwide
More informationDes cellules-souches dans le poumon : pourquoi faire?
Des cellules-souches dans le poumon : pourquoi faire? Karl-Heinz Krause Dept. of Pathology and Immunology, Medical Faculty Dept. of Genetic and Laboratory Medicine, University Hospitals Geneva, Switzerland
More informationANAT 2341 Embryology Lecture 18 Stem Cells
ANAT 2341 Embryology Lecture 18 Stem Cells 29 September 2010 Dr Antonio Lee Neuromuscular & Regenera
More informationSUPPLEMENTARY INFORMATION
DOI: 10.1038/ncb2239 Hepatocytes (Endoderm) Foreskin fibroblasts (Mesoderm) Melanocytes (Ectoderm) +Dox rtta TetO CMV O,S,M,K & N H1 hes H7 hes H9 hes H1 hes-derived EBs H7 hes-derived EBs H9 hes-derived
More informationFundamental properties of Stem Cells
Stem cells Learning Goals: Define what a stem cell is and describe its general properties, using hematopoietic stem cells as an example. Describe to a non-scientist the current progress of human stem cell
More informationDirected Differentiation of Human Induced Pluripotent Stem Cells. into Fallopian Tube Epithelium
Directed Differentiation of Human Induced Pluripotent Stem Cells into Fallopian Tube Epithelium Nur Yucer 1,2, Marie Holzapfel 2, Tilley Jenkins Vogel 2, Lindsay Lenaeus 1, Loren Ornelas 1, Anna Laury
More informationStem Cells: A Primer National Institutes of Health
Stem Cells: A Primer National Institutes of Health I. INTRODUCTION Research on stem cells is advancing knowledge about how an organism develops from a single cell and how healthy cells replace damaged
More informationRegulated Nodal signaling promotes differentiation of the definitive endoderm and mesoderm from ES cells
2078 Research Article Regulated Nodal signaling promotes differentiation of the definitive endoderm and mesoderm from ES cells Masanori Takenaga 1,2, Miki Fukumoto 2 and Yuichi Hori 1,2, * 1 Department
More informationNeural Induction. Chapter One
Neural Induction Chapter One Fertilization Development of the Nervous System Cleavage (Blastula, Gastrula) Neuronal Induction- Neuroblast Formation Cell Migration Mesodermal Induction Lateral Inhibition
More informationXenopus gastrulation. Dorsal-Ventral Patterning - The Spemann Organizer. Two mesoderm inducing signals
Dorsal- atterning - The Spemann Organizer Two mesoderm inducing signals late-blastula early-gastrula nimal Blood Mesothelium Not Vegetal NC Endoderm Hans Spemann (1869-1941) Hilde Mangold (1898-1924) Dr
More informationSingle-cell RNA-seq reveals novel regulators of human embryonic stem cell differentiation to definitive endoderm
Chu et al. Genome Biology (2016) 17:173 DOI 10.1186/s13059-016-1033-x RESEARCH Open Access Single-cell RNA-seq reveals novel regulators of human embryonic stem cell differentiation to definitive endoderm
More informationStem Cell Services. Driving Innovation for Stem Cell Researchers
Driving Innovation for Stem Cell Researchers Stem Cell Services Partner with us and have access to the most advanced and comprehensive stem cell services available today. 675 W. Kendall St. Cambridge,
More informationReading. Lecture III. Nervous System Embryology. Biology. Brain Diseases. September 5, Bio 3411 Lecture III. Nervous System Embryology
Reading NEUROSCIENCE: 5 th ed, pp. 477-506 NEUROSCIENCE: 4 th ed, pp. 545-575 Bio 3411 Wednesday 2 Summary from Lecture II Biology Understanding the brain is THE major question in biology and science.
More informationStem Cells and Cell Culture Developments Within ThermoFisher Scientific
The world leader in serving science Stem Cells and Cell Culture Developments Within ThermoFisher Scientific Katrina Kidd European Application Scientist Cellomics ThermoFisher Scientific A Typical Life
More informationLecture III. Nervous System Embryology
Bio 3411 Wednesday Reading NEUROSCIENCE: 5 th ed, pp. 477-506 NEUROSCIENCE: 4 th ed, pp. 545-575 2 1 Summary from Lecture II Biology Understanding the brain is THE major question in biology and science.
More informationStem Cells for the Future Treatment of Parkinson's Disease
Home Stem Cell Basics Stem Cell Basics I. Introduction II. III. IV. What are the unique properties of all stem cells? What are embryonic stem cells? What are adult stem cells? V. What are the similarities
More informationAt the conclusion of this lesson you should be able to:
Learning Objectives At the conclusion of this lesson you should be able to: Understand the key terms and definitions regarding stem cells Differentiate between the adult and embryonic stem cells Differentiate
More informationWe are walking and standing with parts of our bodies which could have been used for thinking had they developed in another part of the embryo.
We are walking and standing with parts of our bodies which could have been used for thinking had they developed in another part of the embryo. Hans Spemann, 1943 Reading from Chapter 3 - types of cell
More informationSupplementary Figure 1
Supplementary Figure 1 Supplementary Figure 1: Vector maps of TRMPV and TRMPVIR variants. Many derivatives of TRMPV have been generated and tested. Unless otherwise noted, experiments in this paper use
More informationGENESDEV/2007/ Supplementary Figure 1 Elkabetz et al.,
GENESDEV/2007/089581 Supplementary Figure 1 Elkabetz et al., GENESDEV/2007/089581 Supplementary Figure 2 Elkabetz et al., GENESDEV/2007/089581 Supplementary Figure 3 Elkabetz et al., GENESDEV/2007/089581
More information1. Introduction. 1 Page 1
1. INTRODUCTION Osteoblast differentiation is the key component of the bone formation, growth and remodeling. During this incompletely understood process a subpopulation of multipotential mesenchymal stem
More informationABSTRACT INTRODUCTION Figure 1. METHODS ipsc culture ATCC No. Initial Cell Type Description Reprogramming Method Table 1.
THE ESSENTIALS OF LIFE SCIENCE RESEARCH GLOBALLY DELIVERED DIRECTED DIFFERENTIATION OF GASTROINTESTINAL EPITHELIAL ORGANOIDS USING ATCC CELLMATRIX BASEMENT MEMBRANE FROM MULTIPLE HUMAN ATCC ipsc LINES
More informationANAT 3231 Cell Biology Lecture 21 Stem Cells
ANAT 3231 Cell Biology Lecture 21 Stem Cells Outline What are Stem Cells? Totipotency - Pluripotency - Multipotency What are different sources of Stem Cells? Embryonic vs Adult Pros and Cons for each type
More informationCulture and differentiation of embryonic stem cells. Hong-Lin Su Department of Life Sciences, National Chung-Hsing University
Culture and differentiation of embryonic stem cells Hong-Lin Su Department of Life Sciences, National Chung-Hsing University Topics-ES cell maintenance Establishment Culture condition, including the serum,
More informationWhat s the most complex problem in biology?
Chapter 47. Development What s the most complex problem in biology? 1 The most complex problem How to get from here to there Development: cellular level Cell division Differentiation cells become specialized
More informationChapter 47. Development
Chapter 47. Development What s the most complex problem in biology? The most complex problem How to get from here to there Development: cellular level Cell division Differentiation cells become specialized
More informationContents. The Right Surface for Every Cell Extracellular Matrices and Biologically Coated Surfaces ECM Mimetic and Advanced Surfaces...
Contents The Right Surface for Every Cell... 1 Extracellular Matrices and Biologically Coated Surfaces... 2 Corning Matrigel Matrix... 2 Corning BioCoat Cultureware... 3 ECM Mimetic and Advanced Surfaces...
More informationThe Role of the SHB Adapter Protein in Cell Differentiation and Development
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine 150 The Role of the SHB Adapter Protein in Cell Differentiation and Development VITEZSLAV KRIZ ACTA UNIVERSITATIS UPSALIENSIS
More informationSupplemental Figure 1
Supplemental Figure 1 A B 50 * 40 30 20 10 0 NeurofilamentM/βactin Relative expression control D neurogenic cond. PPARγ/ Relative expression C PPARγ Hprt OilRedO adipogenic cond. control 125 * 100 75 50
More informationGenetic Labeling Techniques
Genetic Labeling Techniques CRRT 2012 San Diego, California February 14, 2012 Benjamin D. Humphreys, M.D., Ph.D. Principal Faculty and Co-Director, Harvard Stem Cell Institute Kidney Program Assistant
More informationADVANCED MEDIA TECHNOLOGY
ADVANCED MEDIA TECHNOLOGY For Human ES and ips Cell Research The right medium makes all the difference in ensuring successful ES and ips cell culture. Stemgent offers high-quality, chemically-defined,
More informationSUPPLEMENTARY INFORMATION
a before amputation regeneration regenerated limb DERMIS SKELETON MUSCLE SCHWANN CELLS EPIDERMIS DERMIS SKELETON MUSCLE SCHWANN CELLS EPIDERMIS developmental origin: lateral plate mesoderm presomitic mesoderm
More informationActivity 47.1 What common events occur in the early development of animals? 1. What key events occur at each stage of development?
Notes to Instructors Chapter 47 Animal Development What is the focus of this activity? Chapter 21 provided a review of how genes act to control development. Chapter 47 reviews some of the major morphological
More informationBackground. Background. Background. Background. Background 3/4/2013
3/4/213 Human Very Small Embryonic-like Stem Cells (s) Capacity to Regenerate Bone Tissue: A Potential Cell-based Therapy for Autogenous Bone Grafting Aaron M. Havens, DMD Need to discover an improved
More informationFoxH1 (Fast) functions to specify the anterior primitive streak in the mouse
FoxH1 (Fast) functions to specify the anterior primitive streak in the mouse Pamela A. Hoodless, 1,4 Melanie Pye, 1 Claire Chazaud, 1 Etienne Labbé, 2 Liliana Attisano, 2 Janet Rossant, 1,3 and Jeffrey
More informationImpact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells
JOURNAL OF ANIMAL SCIENCE AND BIOTECHNOLOGY Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells Mohamad-Fauzi et al. Mohamad-Fauzi et al. Journal of Animal
More informationThe Use of Genetically-Modified Mouse Models to Study the Actin Cytoskeleton
The Use of Genetically-Modified Mouse Models to Study the Actin Cytoskeleton Anthony Kee (PhD) Cellular and Genetic Medicine Unit School of Medical Sciences (a.kee@unsw.edu.au) 2017 Structure of the Prac
More informationSolution Key Problem Set
Solution Key- 7.013 Problem Set 5-2013 Question 1 During a summer hike you suddenly spot a huge grizzly bear. This emergency situation triggers a fight or flight response through a signaling pathway as
More informationCharacteristics. Capable of dividing and renewing themselves for long periods of time (proliferation and renewal)
STEM CELLS (p 2-8) Overview The body is made up of about 200 different kinds of specialised cells, such as muscle cells, nerve cells, fat cells and skin cells. Cells with the capacity to give rise to the
More information2/12/2009. Virus enters here. Rakic, JCN, 1972
Stem Cell Update Feb. 12, 2009 Arnold R. Kriegstein, MD, PhD, Department of Neurology, Director, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research at UCSF Virus enters here Rakic,
More informationMouse Engineering Technology. Musculoskeletal Research Center 2016 Summer Educational Series David M. Ornitz Department of Developmental Biology
Mouse Engineering Technology Musculoskeletal Research Center 2016 Summer Educational Series David M. Ornitz Department of Developmental Biology Core service and new technologies Mouse ES core Discussions
More informationEarly Development and Axis Formation in Amphibians
Biology 4361 Early Development and Axis Formation in Amphibians October 25, 2006 Overview Cortical rotation Cleavage Gastrulation Determination the Organizer mesoderm induction Setting up the axes: dorsal/ventral
More informationJohn Gurdon was testing the hypothesis of genomic equivalence or that when cells divide they retain a full genomic compliment.
1. (15 pts) John Gurdon won the 2012 Nobel Prize in Physiology or Medicine for work he did in the 1960 s. What was the major developmental hypothesis he set out to test? What techniques did he development
More informationSupplemental Information. Antagonistic Activities of Sox2. and Brachyury Control the Fate Choice. of Neuro-Mesodermal Progenitors
Developmental Cell, Volume 42 Supplemental Information Antagonistic Activities of Sox2 and Brachyury Control the Fate Choice of Neuro-Mesodermal Progenitors Frederic Koch, Manuela Scholze, Lars Wittler,
More informationDevelopmental Zoology (ZOO ) Gatrulation
Developmental Zoology (ZOO 228.1.0) Gatrulation 1 Developmental Stages Ø Early Development Fertilization Cleavage Gastrulation Neurulation Ø Later Development Organogenesis Larval molts Metamorphosis Aging
More informationNeural Induction. Steven McLoon Department of Neuroscience University of Minnesota
Neural Induction Steven McLoon Department of Neuroscience University of Minnesota 1 Course News Coffee Hour (with Dr. Nakagawa) Friday, Sept 22 8:30-9:30am Surdyks Café in Northrop Auditorium Stop by for
More informationReadings. Lecture IV. Mechanisms of Neural. Neural Development. September 10, Bio 3411 Lecture IV. Mechanisms of Neural Development
Readings Lecture IV. Mechanisms of Neural NEUROSCIENCE: References : 5 th ed, pp 477-506 (sorta) 4 th ed, pp 545-575 (sorta) Fainsod, A., Steinbeisser, H., & De Robertis, E. M. (1994). EMBO J, 13(21),
More information7.22 Example Problems for Exam 1 The exam will be of this format. It will consist of 2-3 sets scenarios.
Massachusetts Institute of Technology Department of Biology 7.22, Fall 2005 - Developmental Biology Instructors: Professor Hazel Sive, Professor Martha Constantine-Paton 1 of 10 7.22 Fall 2005 sample exam
More informationSupplemental Information Inventory
Cell Stem Cell, Volume 6 Supplemental Information Distinct Hematopoietic Stem Cell Subtypes Are Differentially Regulated by TGF-β1 Grant A. Challen, Nathan C. Boles, Stuart M. Chambers, and Margaret A.
More informationTheoretical cloning project
Theoretical cloning project Needed to get credits Make it up yourself, don't copy Possible to do in groups of 2-4 students If you need help or an idea, ask! If you have no idea what to clone, I can give
More informationConcepts and Methods in Developmental Biology
Biology 4361 Developmental Biology Concepts and Methods in Developmental Biology June 16, 2009 Conceptual and Methodological Tools Concepts Genomic equivalence Differential gene expression Differentiation/de-differentiation
More informationBest Practices Maintaining Pluripotency. Anne R. Greenlee, PhD
Best Practices Maintaining Pluripotency Anne R. Greenlee, PhD HESI-Workshop Cary, NC 27 Feb 2007 OHSU La Grande Campus Research Directions Past Fertility Risk Factor Study Pesticide effects on embryo development
More informationInt. J. Mol. Sci. 2016, 17, 1259; doi: /ijms
S1 of S5 Supplementary Materials: Fibroblast-Derived Extracellular Matrix Induces Chondrogenic Differentiation in Human Adipose-Derived Mesenchymal Stromal/Stem Cells in Vitro Kevin Dzobo, Taegyn Turnley,
More informationMouse Mesenchymal Stem Cell Functional Identification Kit
Mouse Mesenchymal Stem Cell Functional Identification Kit Catalog Number SC010 Reagents for the identification of mouse bone marrow-derived stem cells (BMSC)/mesenchymal stem cells (MSC) by in vitro functional
More informationSupplemental Information. Tissue Mechanics Orchestrate Wnt-Dependent. Human Embryonic Stem Cell Differentiation
Cell Stem Cell, Volume 19 Supplemental Information Tissue Mechanics Orchestrate Wnt-Dependent Human Embryonic Stem Cell Differentiation Laralynne Przybyla, Johnathon N. Lakins, and Valerie M. Weaver Supplemental
More informationSupporting Information Lin et al.
Supporting Information Lin et al. SI Materials and Methods. Isolation and culture of human MSCs and ECFCs Human watmscs were isolated from normal discarded subcutaneous white adipose tissue obtained during
More informationThe many faces of Pluripotency: in vitro adaptations of a continuum of in vivo states
Morgani et al. BMC Developmental Biology (2017) 17:7 DOI 10.1186/s12861-017-0150-4 REVIEW The many faces of Pluripotency: in vitro adaptations of a continuum of in vivo states Sophie Morgani 1,2, Jennifer
More informationClip 1: Hip dysplasia Clip 2: Vet-Stem s Dr. Harman
Stem Cells Many breeds of large dogs are affected by hip dysplasia. This degenerative disease affects their hind legs, hindering mobility. Watch as scientists at Vet-Stem explain how stem cells are used
More informationGeneration of Scaffoldless Hyaline Cartilaginous Tissue from Human ipscs
Stem Cell Reports Supplemental Information Generation of Scaffoldless Hyaline Cartilaginous Tissue from Human ipscs Akihiro Yamashita, Miho Morioka, Yasuhito Yahara, Minoru Okada, Tomohito Kobayashi, Shinichi
More informationCycles of vascular plexus formation within the nephrogenic zone of the developing mouse kidney
1 Supplementary text and data for: 2 3 4 5 Cycles of vascular plexus formation within the nephrogenic zone of the developing mouse kidney Authors: David A. D. Munro 1*, Peter Hohenstein 2, and Jamie A.
More informationCell sorting, cell tension and morphogenesis. Morphogenesis: Big Questions
Cell sorting, cell tension and morphogenesis Morphogenesis: Big Questions How are tissues formed from populations of cells? How are organs constructed from tissues? How do organs form in particular locations,
More informationThe Role of Adult Stem Cells in Personalized and Regenerative Medicine
The Role of Adult Stem Cells in Personalized and Regenerative Medicine Christopher J. Neill, Director of Corporate Operations American CryoStem Corporation Objective To illustrate the potential benefits
More informationIntroducing new DNA into the genome requires cloning the donor sequence, delivery of the cloned DNA into the cell, and integration into the genome.
Key Terms Chapter 32: Genetic Engineering Cloning describes propagation of a DNA sequence by incorporating it into a hybrid construct that can be replicated in a host cell. A cloning vector is a plasmid
More informationLife Sciences. Novel ECM Mimetic Surfaces for Cell Expansion and Differentiation
Life Sciences Novel ECM Mimetic Surfaces for Cell Expansion and Differentiation ASCB December 17, 2012 Marshall Kosovsky, Ph.D., Technical Support Manager Topics for Discussion Introduction Overview of
More informationAdvances in understanding tissue regenerative capacity and mechanisms in animals
Advances in understanding tissue regenerative capacity and mechanisms in animals Kenneth D. Poss Abstract Questions about how and why tissue regeneration occurs have captured the attention of countless
More informationA novel stem cell for auditory neuron regeneration.
A novel stem cell for auditory neuron regeneration. M.A.Huisman*, R. el Seady*, C.F.H.M.Löwik # en J.H.M.Frijns* Departments of Ear, Nose & Throat* and Molecular Imaging #, Leiden University Medical Centre,
More informationV4: cellular reprogramming
V4: cellular reprogramming Embryonic development Stem cells Differentiation ips cells Oct4 Paper4 Model of a human cell, Spiegel Online 3D-Model of the human genome, Spiegel Online Some human cells Astrocyte
More informationSupplemental Information
Supplemental Information Itemized List Materials and Methods, Related to Supplemental Figures S5A-C and S6. Supplemental Figure S1, Related to Figures 1 and 2. Supplemental Figure S2, Related to Figure
More informationHuman Adipose Tissue Is a Source of Multipotent Stem Cells
Molecular Biology of the Cell Vol. 13, 4279 4295, December 2002 Human Adipose Tissue Is a Source of Multipotent Stem Cells D Patricia A. Zuk,* Min Zhu,* Peter Ashjian,* Daniel A. De Ugarte,* Jerry I. Huang,*
More informationFertilization. Animal hemisphere. Sperm entry point
Fertilization Animal hemisphere Sperm entry point Establishes the dorsal/ventral axis Ventral side sperm entry Dorsal side gray crescent Organized by sperm centriole Cleavage Unequal radial holoblastic
More informationIsolation and Analysis of Pluripotent, Neural, and Hematopoietic Stem Cells
Isolation and Analysis of Pluripotent, Neural, and Hematopoietic Stem Cells Christian Carson BD Biosciences R&D Scientist Stem Cell 23-10679-00 Overview Introduction Challenges in stem cell research Antibody
More informationNucleofector Technology in. Somatic Stem Cell Research. gene transfer begins here
Nucleofector Technology in Somatic Stem Cell Research The Nucleofector technology The first highly efficient method for non-viral gene transfer into primary cells and difficult-to-transfect cell lines
More informationSUPPLEMENTARY INFORMATION
doi:10.1038/nature10163 Supplementary Table 1 Efficiency of vector construction. Process wells recovered efficiency (%) Recombineering* 480 461 96 Intermediate plasmids 461 381 83 Recombineering efficiency
More informationPPAR-γ - Peroxisome proliferator-ac4vated receptor gamma
PPAR-γ - Peroxisome proliferator-ac4vated receptor gamma PPAR-γ - Peroxisome proliferator-ac4vated receptor gamma C/EBPalpha - CCAAT/enhancer-binding protein alpha C/EBPα contains: - a func)onally related
More informationRole of Bmi-1 in Epigenetic Regulation. During Early Neural Crest Development
Role of Bmi-1 in Epigenetic Regulation During Early Neural Crest Development Thesis by Jane I. Khudyakov In Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy California Institute
More informationINVESTIGATION OF MSC DIFFERENTIATION ON ELECTROSPUN NANOFIBROUS SCAFFOLDS
With support of NSF Award no. EEC-0754741 INVESTIGATION OF MSC DIFFERENTIATION ON ELECTROSPUN NANOFIBROUS SCAFFOLDS NSF Summer Undergraduate Fellowship in Sensor Technologies Emily Wible (Bioengineering)
More informationStem Cells: Ethics and Oversight May 24, Sidney H. Golub, Ph.D. Interim Director Sue & Bill Gross Stem Cell Research Center
Stem Cells: Ethics and Oversight May 24, 2017 Sidney H. Golub, Ph.D. Interim Director Sue & Bill Gross Stem Cell Research Center Nobel Laureates 2012 John Gurdon Shinya Yamanaka Pluripotent Stem Cell PLURIPOTENT
More informationHUMAN ADIPOSE TISSUE IS A SOURCE OF MULTIPOTENT STEM CELLS. Patricia A. Zuk, PhD* Min Zhu, MD* Peter Ashjian, MD* Daniel A.
MBC in Press, published on September 24, 2002 as 10.1091/mbc.E02-02-0105 HUMAN ADIPOSE TISSUE IS A SOURCE OF MULTIPOTENT STEM CELLS Patricia A. Zuk, PhD* Min Zhu, MD* Peter Ashjian, MD* Daniel A. De Ugarte,
More informationNanoparticle labeling identifies slow cycling human endometrial stromal cells
Xiang et al. Stem Cell Research & Therapy 2014, 5:84 RESEARCH Nanoparticle labeling identifies slow cycling human endometrial stromal cells Lina Xiang 1, Rachel W S Chan 1,2*, Ernest H Y Ng 1,2 and William
More informationIsolation and biological characterization of chicken amnion epithelial cells
European Journal of Histochemistry 2012; volume 56:e33 Isolation and biological characterization of chicken amnion Y. Gao, 1 Y. Pu, 1 D. Wang, 2 W. Zhang, 1 W. Guan, 1 Y. Ma 1 1 Institute of Animal Sciences,
More information2. Outline the levels of DNA packing in the eukaryotic nucleus below next to the diagram provided.
AP Biology Reading Packet 6- Molecular Genetics Part 2 Name Chapter 19: Eukaryotic Genomes 1. Define the following terms: a. Euchromatin b. Heterochromatin c. Nucleosome 2. Outline the levels of DNA packing
More informationNatural history of mesenchymal stem cells, from vessel walls to culture vessels
Cell. Mol. Life Sci. DOI 10.1007/s00018-013-1462-6 Cellular and Molecular Life Sciences REVIEW Natural history of mesenchymal stem cells, from vessel walls to culture vessels Iain R. Murray Christopher
More informationDifferentiation of Embryonic Stem Cells to Clinically Relevant Populations: Lessons from Embryonic Development
Leading Edge Review Differentiation of Embryonic Stem Cells to Clinically Relevant Populations: Lessons from Embryonic Development Charles E. Murry 1 and Gordon Keller 2, * 1 Departments of Pathology and
More informationTime allowed: 2 hours Answer ALL questions in Section A, ALL PARTS of the question in Section B and ONE question from Section C.
UNIVERSITY OF EAST ANGLIA School of Biological Sciences Main Series UG Examination 2013-2014 CELL BIOLOGY BIO-2B06 Time allowed: 2 hours Answer ALL questions in Section A, ALL PARTS of the question in
More informationPeter Dy, Weihuan Wang, Pallavi Bhattaram, Qiuqing Wang, Lai Wang, R. Tracy Ballock, and Véronique Lefebvre
Developmental Cell, Volume 22 Supplemental Information Sox9 Directs Hypertrophic Maturation and Blocks Osteoblast Differentiation of Growth Plate Chondrocytes Peter Dy, Weihuan Wang, Pallavi Bhattaram,
More informationGuide to Clonetics and Poietics Cell Types
Guide to Clonetics and Poietics Cell Types Clonetics and Poietics Cell Types - Sorted By Tissue Source Cryopreserved Human Cells Adipose Subcutaneous Preadipocytes PT-5001 PGM -2 PT-8002 Visceral Preadipocytes
More informationHuman embryonic stem cells for in-vitro developmental toxicity testing
Human embryonic stem cells for in-vitro developmental toxicity testing HESI workshop on alternatives assays for developmental toxicity Raimund Strehl Cellartis The company Founded in early 2001, University
More informationCharles Shuler, Ph.D. University of Southern California Los Angeles, California Approved for Public Release; Distribution Unlimited
AD Award Number: DAMD17-01-1-0100 TITLE: Smad-Mediated Signaling During Prostate Growth and Development PRINCIPAL INVESTIGATOR: Charles Shuler, Ph.D. CONTRACTING ORGANIZATION: University of Southern California
More informationSUPPLEMENTARY INFORMATION
doi:10.1038/nature11435 Supplementary Figures Figure S1. The scheme of androgenetic haploid embryonic stem (ahes) cells derivation. WWW.NATURE.COM/NATURE 1 RESEARCH SUPPLEMENTARY INFORMATION Figure S2.
More informationEts-1 Confers Cranial Features on Neural Crest Delamination
Ets-1 Confers Cranial Features on Neural Crest Delamination Eric Théveneau 1,2, Jean-Loup Duband 1,2, Muriel Altabef 1,2 * 1 Université Pierre et Marie Curie - Paris6, UMR 7622, Laboratoire de Biologie
More information