UNSW Embryology

 GENM0202 - Lecture - Neural Stem Cells

© Dr Mark Hill (2009)

Acknowledgements

Introduction

	This GENM0202 lecture is an introduction to stem cell applications in neuroscience. The lecture will introduce the concept of a "Stem Cell" and their various origins. Then I will focus on the applications, both current and potential, of stem cells in neuroscience. More information can be found in Neural Stem Cell Notes
Cellular components of the central nervous system	Some content in this current lecture is from the NIH Publications: "Regenerative Medicine" Department of Health and Human Services (August 2006) and "Stem Cells: Scientific Progress and Future Research Directions" (2001) and

Page Links: Introduction | Lecture Slides | Lectopia Audio | Lecture Objectives | UNSW Embryology | Comments | Online Books | Internet Links | Glossary | Terms |

Lecture Slides

These links are to PDF versions of the Lecture slides: 1 slide/page viewing 48 pages | 3 slide/page printing 12 pages | 6 slide/page printing 8 pages

Lectopia Audio Link to the audio recording of this lecture will be added here when available.

The online text Online Books and UNSW Embryology links listed on this page worth reviewing for this lecture.

Lecture Objectives

• Understanding of stem cell definition
• Understanding of stem cell sources
• Understanding of neural stem cells
• Brief understanding of neural stem cell applications

Lecture Summary

The following text is extracted and modified from the previous 2006 lecture slides and should be used as a "trigger" to remind you of key concepts.

• Background to Stem Cells
  • Medline Papers Search
  • “Stem cell” 110,920 (2002), 127,869 (2003), 128,485 (2004), 140,966 (2005), 156,015 (2006), 212,765 (2008)
  • “Stem cells” 102,324 (2002), 115,680 (2003), 116,229 (2004), 125,722 (2005), 137,921 (2006), 189,713 (2008)
  • NIH Stem Cells http://stemcells.nih.gov/index.asp
  • Stem Cell Basics http://stemcells.nih.gov/info/basics/
  • Stem Cells Report (2001) http://stemcells.nih.gov/info/scireport/
• Stem Cells
• Establishment of order from 1 cell
• Clonal, stem cells
• Stem Cells in the News- Australia
  • Abbott denies stem cell report is skewed
    Health Minister Tony Abbott has denied the government bought an opinion on stem cell research in an attempt to skew the debate on therapeutic cloning. 2008-09-01
    Lib anger over PM's secret stem cell report
    A REPORT commissioned by the Prime Minister challenging the findings of an expert government review on embryo research has drawn sharp criticism from his own MPs.2008-09-01
    Govt bought stem cell report: Labor
    Labor has attacked the independence of a new report on stem cell research. 2008-09-01
    New cloning report sparks controversy
    A new report which could undermine the push to overturn a ban on therapeutic cloning has been released by Prime Minister John Howard ahead of a conscience vote on the issue. 2008-08-31
    PM criticised over stem cell debate
    Two prominent Liberal MPs have criticised Prime Minister John Howard's latest intervention in the stem cell debate, accusing him of shopping around for the advice he wants to hear. 2008-08-31
• Stem Cells in the News - World
  • Stem cell treatment for ill girl
    Umbilical cord stem cells are being used for Sacha which would normally be incinerated.” 28 Aug 2008
    'Ethical' stem cell lines created
    Of the 16 embryos used, they developed two long-term stem cell lines, which, Professor Lanza said, were "genetically normal and able to generate all of the cell types of the body". 24 Aug 2008
    Bush uses veto on stem cell bill
    The stem cell debate seems set to be an issue in November's mid-term congressional elections. 19 Jul 2008
    US Senate approves stem cell bill
    One encourages stem cell research using cells from sources other than embryos, and the other bill bans the growing and aborting of foetuses for research. 18 Jul 2008
    Euro MPs debate stem cell funding
    Only a tiny proportion of EU science funding for 2007-13 will go to stem cell research, under current plans. Only three countries, Britain, Sweden and Belgium, allow research using cloned stem cells. 13 Jun 2008
• 3 Adult Cell Types (Proliferation)
  
• Differentiated cells no longer capable of cell division
  Cardiac muscle cells, neurons
  produced during embryonic development, differentiate, and are then retained throughout the life of the organism.
  Cells in G0 stage of cell cycle
  resume proliferation when needed to replace cells
  skin fibroblasts, smooth muscle cells, endothelial cells, epithelial cells of internal organs, such as the liver, pancreas, kidney, lung, prostate and breast
• Stem Cells
  differentiated cells with short life spans continually replaced
  blood cells, epithelial cells of skin and digestive tract
  fully differentiated cells do not themselves proliferate
  proliferation of cells that are less differentiated- stem cells
  divide to produce daughter cells that either differentiate or remain as stem cells
• Blood Cells
  All different types of blood cells develop from a pluripotent stem cell in bone marrow
  Precursors of differentiated cells undergo several rounds of cell division as they mature
  proliferation ceases at terminal stages of differentiation
  Movie: Neural Stem Cell
• Research that led to Stem Cells
  • Human Diseases
    Generation of “knock out” mice
    Human Development
    Studying regulation of cell differentiation in development
    Human Reproduction
    Disorders, sterility
• Stem cells in Development
  Blastocyst and Cord blood
  
• Pluripotent Stem Cells
  Pluripotent
  to describe stem cells that can give rise to cells derived from all 3 embryonic germ layers
  Mesoderm, Endoderm, Ectoderm
  layers are embryonic source of all cells of the body
  Mouse ES Cells
  Blastocyst
  hollow structure composed of about 100 cells surrounding an inner cavity
  Only ES cells, which form inner cell mass, actually form the embryo.
  ES cells can be removed from the blastocyst and grown on lethally irradiated “feeder cells.”
  See E. Robertson et al., 1986, Nature 323:445.
  
• What is a stem cell- Definition
  • Stem cell is a cell that has the ability to divide (self replicate) for indefinite periods throughout life of organism
  • Under the right conditions, or given the right signals, stem cells can differentiate to the many different cell types that make up the organism
    
• Chimeric Mouse
  Generated with ES or teratocarcinoma stem cells
  shows that stem cells can combine with the cells of a normal blastocyst to form a healthy chimeric mouse
  
• Historic References
  
  • Mouse
    • Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Martin GR. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7634-8.
      
    • Characterization of a pluripotent stem cell line derived from a mouse embryo. Wobus AM, Holzhausen H, Jakel P, Schoneich J. Exp Cell Res. 1984 May;152(1):212-9.
    • Transgenesis by means of blastocyst-derived embryonic stem cell lines. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9065-9. Gossler A, Doetschman T, Korn R, Serfling E, Kemler R.
  • Pig and Sheep
    • Derivation of pluripotent, embryonic cell lines from the pig and sheep. Notarianni E, Galli C, Laurie S, Moor RM, Evans MJ. J Reprod Fertil Suppl. 1991;43:255-60.
      
  • Primate
    
    • Isolation of a primate embryonic stem cell line. Thomson JA, Kalishman J, Golos TG, Durning M, Harris CP, Becker RA, Hearn JP. Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7844-8.
      
  • Human
    • Embryonic stem cell lines derived from human blastocysts.
      Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS, Jones JM. Science. 1998 Nov 6;282(5391):1145-7.
• Why are they in the News?
  Scientific and Ethical
  Therapeutic uses
  Issues relating to human cloning
  Use of excess human eggs/sperm for research purposes
  Availability of human stem cell lines
  What can they be used for?
  Generation of “knock out” mice
  Studying regulation of cell differentiation in development
  Therapeutic uses?
  Genetic disease
  
• Neurodegenerative
  Injury
  Cultivation
  Four?Human?ES?Cell Lines
  Long Term ES Cell Culture
  Human ES Cell Differentiation
  
• Stem Cells in the Adult
  
  • Connective Tissue
    
  • Bone marrow
    
  • Blood Cells, Osteoclasts, blasts
    
  • Epithelia
    
  • Gut
    
  • Skin
  • Epidermis: Immortal Stem Cell
    Each self-renewing patch of epidermis
    contain in each generation at least one "immortal" stem cell
    whose descendants will still be present in the patch in the distant future
    Other basal cells
    born chemically different in a way that commits to leave basal layer and differentiate
    or might be stem cells, equivalent to immortal stem cell in character and mortal only in sense that their progeny happen subsequently to be jostled out of the basal layer and shed from the skin
    Amplifying Cells
    Stem cells in many tissues divide only rarely
    give rise to transit amplifying cells
    daughters committed to differentiation that go through a limited series of more rapid divisions before completing the process.
    each stem cell division gives rise in this way to eight terminally differentiated progeny
    (Ab)Normal Stem Cell Production
    (A) normal strategy for producing new differentiated cells
    (B and C) 2 types of derangement that can give rise to unbridled proliferation characteristic of cancer
    
• Stem Cell Daughter Fates
  
  • environmental asymmetry
    
  • daughters are initially similar
    
  • directed into different pathways according to environmental influences that act on them after they are born
    
  • number of stem cells can be increased or reduced to fit niche available
    
  • divisional asymmetry
    
  • stem cell has an internal asymmetry
    
  • divides in such a way that its two daughters are already endowed with different determinants at time of their birth
    
• Neural Stem Cells: Development
  
• Neural Stem Cells: Adult
  
• Current research on stem cells
  
  • How to: Isolate, Grow, Maintain, store, Differentiate and Therapeutic uses
    
• Stem Cell Markers
  Surface of every cell have specialized proteins (receptors) that have the capability of selectively binding or adhering to other “signalling” molecules
  Many different types of receptors that differ in their structure and affinity for signalling molecules
  Cells use receptors and molecules that bind to them as a way of communicating with other cells
  These same cell surface receptors are stem cell markers
  Stage-Specific Embryonic Antigen-1,4
  Tumor Rejection Antigen (TRA-1-60)
  Stem Cell Antigen 1 (Sca-1)
  
• ES Therapeutics
  
  • Neural
    Parkinson’s, ALS, spinal cord injury……..
    Cell Replacement
    cell death, loss of function
    Grafting
    where host-graft rejection normally requires substantial ongoing immunosuppression
    Repair
    Spinal cord and brain injury
    Other Diseases
    Diabetes, muscular dystrophies, cardiac, vital organs……
• Recent References
  • Neural precursors derived from human embryonic stem cells maintain long-term proliferation without losing the potential to differentiate into all three neural lineages, including dopaminergic neurons. Hong S, Kang UJ, Isacson O, Kim KS. J Neurochem. 2008 Jan;104(2):316-24.
  • Migration of transplanted neural progenitor cells in a ferret model of cortical dysplasia. Schaefer AW, Juliano SL. Exp Neurol. 2008 Mar;210(1):67-82. Epub 2007 Oct 23.
  • Adult olfactory bulb neural precursor cell grafts provide temporary protection from motor neuron degeneration, improve motor function, and extend survival in amyotrophic lateral sclerosis mice. Martin LJ, Liu Z. J Neuropathol Exp Neurol. 2007 Nov;66(11):1002-18.
  • Behavioral, temporal, and spatial targets for cellular transplants as adjuncts to rehabilitation for stroke. Dobkin BH. Stroke. 2007 Feb;38(2 Suppl):832-9. Review
  • Isolation of neural stem cells from the postnatal cerebellum. Lee A, Kessler JD, Read TA, Kaiser C, Corbeil D, Huttner WB, Johnson JE, Wechsler-Reya RJ. Nat Neurosci. 2005 Jun;8(6):723-9. Epub 2005 May 22.
  • Human neural stem cell transplants improve motor function in a rat model of Huntington's disease. McBride JL, etal., J Comp Neurol. 2004 Jul 19;475(2):211-9.
  • Rapid neural differentiation of human cord blood-derived mesenchymal stem cells. Jeong JA etal Neuroreport. 2004;15(11):1731-1734.
  • Cell fusion-independent differentiation of neural stem cells to the endothelial lineage. Wurmser AE, etal. Nature. 2004; 430(6997):350-6.

UNSW Embryology

The following links are to UNSW Embryology additional resources that provide further background information on the Lecture topics. Note that not all information found on these additional links is considered examinable and the lecture slides and laboratory classes should be used as your initial guide for course theory content. Links: Introduction to Stem cells | Stem Cell Ethics | Cord Blood | Adult Stem Cells | Neural Stem Cells The search window below allows a search within the UNSW Embryology website.

Coordinator Comments

Dr Mark Hill

This lecture page is being updated for the current course, so it is worth coming back again later to see if any changes have occurred. Please let me know by email of broken links or content that is not clearly covered in this supporting online material.

Online Books

Neuroscience

Generation of Neurons in the Adult Brain  | Parkinson's Disease: An Opportunity for Novel Therapeutic Approaches |
The Initial Formation of the Nervous System: Gastrulation and Neurulation | Neuroglial Cells | Dividing precursor cells in the Initial Differentiation of Neurons and Glia 

Basic Neurochemistry

Central nervous system stem cells provide an opportunity to improve the prospects for successful transplantation | Growth factors regulate oligodendrocyte development | Neuropoietic model of neural crest... (image) | General Development of the Nervous System  | Bypassing the Barriers with Drugs 

Parkinson's Disease

Neural Stem Cell Biology 

Challenges of Neural Stem Cell Transplantation for Parkinson's Disease and Other Neurodegenerative Diseases 

Neural Stem Cells for Transplantation 

Neural Stem Cells 

Possible sources of neural progenitor... (image) 

Summary: Prospects for New Cell Sources 

Comparison of in Vivo and ex Vivo Gene Transfer Approaches 

Cell Vehicles for ex Vivo Gene Transfer for Parkinson's Disease 

Developmental Biology 6th ed. Gilbert, Scott F. Sunderland (MA): Sinauer Associates, Inc.; c2000.

Below are listed links that relate to this Lecture from the textbook "Developmental Biology" which is available free online. You can either click the provided links or do your own search using the search link. Adult neural stem cells 

Human Somatic and Germ Line Gene Therapy 

Embryonic germ (EG) cells 

Germ cell migration in mammals 

ES cell therapeutics. (A) Human Somatic and Germ Line Gene Therapy
Tissue Architecture of the Central Nervous System 

Cerebral organization 

The RTK pathway 

Cutaneous appendages 

Snapshot Summary: Central Nervous System and Epidermis 

Neural regeneration 

Evidence of adult neural stem... 

Fetal Neurons in Adult Hosts 

The mechanisms of trunk neural crest migration 

The pluripotency of trunk neural crest cells. 

Recognition of surrounding extracellular matrices. 

Search Developmental Biology

Molecular Biology of the Cell 4th ed. Alberts, Bruce; Bray, Dennis; Lewis, Julian; Raff, Martin; Roberts, Keith; Watson, James D. New York and London: Garland Publishing; c2002.

Below are listed links that relate to this Lecture from the textbook "Molecular Biology of the Cell" which is available free online. You can either click the provided links or do your own search using the search link.

The Stem Cells of Adult Tissues May Be More Versatile Than They Seem 

Neural Stem Cells Can Repopulate the Central Nervous System 

Lateral Inhibition Drives the Progeny of the Sensory Mother Cell Toward Different Final Fates 

Stem-Cell Engineering Summary 

Localized Expression of Specific Classes of Gene Regulatory Proteins Foreshadows Cell Differentiation 

Programs That Are Intrinsic to a Cell Often Define the Time-Course of its Development 

Search Molecular Biology of the Cell

Internet Links

Journals: Cell - Stem Cells | Stem Cells

Glossary

Terms

Alzheimer's Disease - (AD) A neurodegenerative disease associated mainly with ageing and the most common form of dementia. Involves neurons in the parts of the brain that control thought, memory and language. Histopathology is shown postmortem by extracellular beta-amyloid (Aß) fibrils plus intraneuronal tau (a microtubule associated protein) filaments. Named after Aloysius Alzheimer (1864-1915), a German psychiatrist and neuropathologist first published case of "presenile dementia". (More? Neural Notes | Medline Plus - Alzheimer's Disease)

Amyotrophic Lateral Sclerosis - (ALS, Lou Gehrig's disease) A postnatal neurological disease of the central nervous system, affecting the spinal cord motor neurons and brain. Henry Louis "Lou" Gehrig (1903–1941), was an American baseball player in the 1920s and 1930s who suffered from this disease. (More? Neural Notes | Medline Plus - Amyotrophic Lateral Sclerosis)

blastocyst - (Greek, blastos = sprout + cystos = cavity) The early stage in embryo development, in humans week 1 and 2, before implantation. The blastocyst consists of cells forming an outer trophoblast layer, an inner cell mass and a fluid-filled cavity. The blastocyst inner cell mass is the source of true embryonic stem cells capable of forming all cell types within the embryo. (More? Week 1 - Blastocyst | Week 2 Notes | Stem Cell Notes)

bone marrow - The cellular components found within the core of bones, mainly long bones, which contain the adult blood stem cell population and a range of other cell types. (More? Musculoskeletal Development - Bone | Musculoskeletal Development)

bone marrow stromal cells - (BMSCs) These are pluripotental cells from bone marrow that can potentially differentiate into a range of connective tissue and muscle cell types (cardiomyocytes, rhabdomyocytes, hepatocytes, osteocytes, chondrocytes, tencoytes, adipocytes, smooth muscle cells). These cells have therefore been seen as a source of stem cells for tissue repair. (More? Musculoskeletal Development - Bone | Musculoskeletal Development)

chimera - (Greek, pronounced ky-MIR-uhs) A mythical monster with the head of a lion, the body of a goat, and the tail of a serpent. Today used to describe a mixing of biological molecules either: 1. animals that have been genetically engineered 2. development studies where different species cells have been mixed, example chick-quail chimera, (More? Neural Crest Notes) 3. Modified proteins containing parts of 2 different proteins.

cord blood - A term used to describe blood collected from the placenta usually after birth. Has been identified as a source of stem cells with potential therapeutic uses and is stored in Cord Blood Banks throughout the world. (More? Placenta Development | Stem Cell)

ES cell - An Embryonic Stem cell derived from the inner cell mass of the blastocyst, which is totipotential and can be grown undifferentiated in tissue culture (in vitro). (More? Stem Cell Notes)

glial cell - A cell within the nervous system that does not itself transmit electrical and chemical signals, but which provides metabolic and structural support for neurons. (More? Neural Notes)

“knock out” mice - A term used to describe the molecular biology technique of introducing a germ line gene specific disruption that results in mice that do not produce the protein product of that gene, it has been "knocked out". Technique required the development of stem cells to carry out the manipulation. (More? Stem Cell Notes | Molecular Notes | Mouse Notes)

medulloblast - An undifferentiated cell of the embryonic neural tube that can develop into either a neuroblast or spongioblast similar to a neural stem cell. A medulloblastoma is a tumor that has been suggested to form from transformed neural stem cells.

medulloblastoma - A tumor that has been suggested to form from transformed neural stem cells.

neural crest - cell region at edge of neural plate, then atop the neural folds, that remains outside and initially dorsal to the neural tube when it forms. These paired dorsal lateral streaks of cells migrate throughout the embryo and can differentiate into many different cell types (= pluripotential). Those that remain on the dorsal neural tube form the sensory spinal ganglia (DRG). Neural crest cells also migrate into the somites. (More? Neural Crest Notes)

neural stem cell (NSC) Stem cells within the central nervous system (CNS) that proliferate indefinitely and give rise to either neurons, astrocytes, and oligodendrocytes.

neural stem cell niche - Locations within the nervous system that multipotent stem cells reside and participate in specialized microenvironments that support self-renewal and differentiation.

neural tube - The third stage in early development of the central nervous system. In the trilaminar embryo (ectoderm, mesoderm, endoderm) the central region of the ectoderm (in the midline above the mesodermal notochord) initially forms a columnar epithelium described as the neural plate. This epithelium will fold dorsally to form the neural groove, which then fuses to form an initially open at either end hollow neural tube. The neural tube forms the central nervous system (brain and spinal cord). Developmental sequence: neural plate -> neural groove -> neural tube -> Central Nervous System -> brain and spinal cord. (More? Neural Notes)

neurogenesis - the process of formation of the neural system. This begins with segregation of the neural plate from the ectoderm of the trilaminar embryo by folding to form initially the neural groove, which then fuses to form the neural tube (the central nervous system progenitor, brain and spinal cord) and associated neural crest. (More? Neural Notes)

neuron - (neurone) The cell forming the unit basis of the nervous system (both central and peripheral) capable of generating an action potential and releasing neurotransmitter. There are many different types of neuronal cells. (More? Neural Notes)

olfactory epithelium - The specialised sensory epithelium that lines the nasal cavity associated with smell. Some cells within the adult epithelium remain as stem cells which can be isolated and purified. (More? Senses Smell | Senses Notes | Stem Cell Notes)

Parkinson's disease - (paralysis agitans, shaking palsy) A postnatal neurological disease of the central nervous system, typically in ageing and more common in men than in women. The neurons affected are those in the brain that control muscle movement and have dopamine as their neurotransmitter. Named after James Parkinson (1730-1813), an English physician who made the detailed description of the disease in "An Essay on the Shaking Palsy" (1817) (More? Medline Plus - Parkinson's Disease) | Pearce JM. Aspects of the history of Parkinson's disease. J Neurol Neurosurg Psychiatry. 1989 Jun;Suppl:6-10)

pluripotent A stem cell term describing the ability of a stem cell to form (differentiate) into many different cell types. The term totipotent means that the stem cell can form all embryo cell types. (More? Stem Cell Notes)

spinal cord - The caudal end of neural tube that, together with the brain (rostral end of neural tube), forms the central nervous system (CNS). Note: the process of secondary neuralation contributes the very caudal end of the spinal cord. (More? Neural Notes)

Stage-Specific Embryonic Antigen-1 - (SSEA-1) A cell surface embryonic marker (antigen) which has a role in cell adhesion, migration and differentiation and is often differentially expressed during stem cell development. Can be identified by Davor Solter monoclonal antibody MC-480 (SSEA-1). (More? Stem Cell Notes)

Stage-Specific Embryonic Antigen 4 - (SSEA-4) A cell surface embryonic marker (antigen) of human teratocarcinoma stem cells (EC), human embryonic germ cells (EG) and human embryonic stem cells (ES) which is down-regulated following differentiation of human EC cells. This antigen is not expressed on undifferentiated murine EC, ES and EG cells but upregulated on differentiation of murine EC and ES cells. Can be identified by Davor Solter monoclonal antibody MC-813-70 (SSEA-4). (More? Stem Cell Notes)

stem cell - Term used to describe a cell with the potential to reproduce itself indefinitely, as well as differentiate into any other embryo tissue cell types. There are also a number of different specialised stem cell definitions: totipotential stem cell (as described above), pluripotential stem cell (capable of forming a number of different cell types), embryonic stem cell (derived from the blastocyst), cord blood stem cell (derived from placental cord blood), and adult stem cell (derived from adult or postnatal tissue). (More? Stem Cell Notes)

Stem Cell Antigen 1 - (Sca-1) A cell surface marker (antigen) for mouse hematopoietic progenitor/stem cell subset. It is a member of the Ly-6 family of GPI-linked surface proteins (Mr 18 kDa). (More? Stem Cell Notes)

totipotent - Term that describes the ability of a cell (stem cell) to form an entire organism or any tissue from that organism. Totipotential suggests a wider differentiation ability than pluripotential, where there is a restriction to a particular group of cell types such as that seen for bone marrow blood stem cells. (More? Stem Cell Notes)

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