Kuldip S Sidhu
A/Professor Kuldip Sidhu, PhD, BSC (Medical) is the chair of stem cell biology and is the director of the Stem Cell Laboratory (SCL), Faculty of Medicine, the University of New South Wales, Australia. His post doc training is from St Louis MO USA (1979-80) in assisted reproductive technology and he also worked with Prof James Thompson, Wisconsin USA (2000) who produced the first human embryonic stem cells in 1998.
His research focus is on neural stem cells derived from both the embryonic and non-embryonic sources for eveloping future cell therapies for various neurodegenerative diseases, like Alzheimer’s, Parkinson’s and other neuronal diseases.
He has established a state-of-the-art facility to study the cell and developmental biology of stem cells. SCL has expertise to culture, propagate, differentiate, engineer and transplant in animal models the neural stem cells from various sources like human embryonic stem cells, skin-derived neuroprogenitors and human mesenchymal stem cells from bone marrow. In addition, he has expertise in the derivation of new human embryonic stem cell lines including their clonal propagation. His lab was first to produce two hESC lines, Endeavour (E) 1&2 from Australia and E2 is listed on NIH registry USA for distribution. SCL has seriously embarked on iPS Technology and produced over 100 iPSC clones from Alzheimer’s patients for studying the disease processes and drug discovery.
A/Prof Sidhu has produced two books – the latest one (2012) on stem cells, thirteen book chapters, nine review chapters, two international patents, four proprietary items and one hundred and seventy original research papers including abstracts in journal of repute including one in Nature Biotechnology (2011) and all dealing with mammalian cell and developmental biology including stem cells. He has served on the International Society of Stem Cell Research Sub Committee and the NHMRC Cell Therapy Advisory Committee; he is a member of the editorial board of International Stem Cell Journals, the open stem cell journal and recent patent on regenerative medicine. He is on the expert panel on iPSC research for the European Union. He is president of the local chapter and member of the board of Society for Brain Mapping and Therapeutics, USA. He was the chair of the program committee of the 4th annual meeting of the Australasian society of stem cell research held in Sydney 2011. He has eight national and five international active research collaborations and including three with industry. He has widely travelled around the world and presented invited lectures, chaired sessions in scientific meetings, conferences.
He is recognised by many awards e.g. Medallist for outstanding research from Indian National Science Academy 1981, Best book prize, 1996, Medal for best presentation in an international conference on frontiers of reproductive biology, 1989, Best invention prize, Australia, 2007, Finalist of Eureka prize 2009, Advanced Innovation Award (Finalist), UNSW 2012. He has produced 9 PhD, 2 MSC, 4 HONS students and some of them are also recognised with Dean’s list and McConaghy Prize.
His passion in science is as good as in Tennis.
Textbooks
Objectives
- Understanding of stem cell history
- Understanding of stem cell types
- Understanding of stem cell identification/differentiation
- Understanding of advantages and disadvantages of different stem cell types
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Lectopia Lecture Audio
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Why Stem Cells
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
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What are their uses?
- Generation of “knock out” mice
- Studying regulation of cell differentiation in development
- Therapeutic uses?
- Genetic disease
- Neurodegenerative
- Injury
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Medline Search stem cell 2002 - 110,920 | 2004 - 128,485 | 2005 - 140,966 | 2006 - 154,176 | 2011 - 190,069 | 2012 - 210,393
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 Cell Types
Tissue Stem Cells
- differentiated cells have short life spans continually replaced
- blood cells, epithelial cells of skin and digestive tract
- fully differentiated cells do not proliferate
- proliferation of less differentiated- stem cells
- produce daughter cells that either differentiate or remain as stem cells
Blood Cells
 Hematopoietic and stromal cell differentiation - 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
Embryonic Stem Cells
NIH - What are embryonic stem cells?
- What is a stem cell - Pluripotent (totipotent)
- Pluripotent - to describe stem cells that can give rise to cells derived from all 3 embryonic germ layers (Ectoderm, Mesoderm, Endoderm)
- layers are embryonic source of all cells of the body
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)
Stem Cell Definition
- cell that has ability to divide for indefinite periods
- self replicate
- throughout life of organism
- stem cells can differentiate
- to the many different cell types
Chimeric Mouse
- ES or teratocarcinoma - shows that stem cells can combine with cells of a normal blastocyst to form a healthy chimeric mouse
Embryoid Bodies
- spheroid cellular tissue culture structure
- mouse and human ES cells have the capacity to undergo controlled differentiation
- recapitulate some aspects of early development
- regional-specific differentiation program
- derivatives of all three embryonic germ layers
Historic References
Mouse
<pubmed>6950406</pubmed>
<pubmed>6714319</pubmed>
<pubmed>3024164</pubmed>
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Pig and Sheep
<pubmed>1843344</pubmed>
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Primate
<pubmed>7544005</pubmed>
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Human
<pubmed>9804556</pubmed>
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Stem Cell Lines ATCC - Embryonic Stem cell lines
Cord Blood Stem Cells
 Cord blood induced stem cell differentiation - Blood collected from the placental umbilical cord of a newborn baby shortly after birth (about 90 ml)
- blood stem cells that can be used to generate red blood cells and cells of the immune system
- collected, typed, stored in Cord Blood Bank (public and private Banks have arisen)
- available for use by the donor and compatible siblings
- suggested use to treat a range of blood disorders and immune system conditions such as leukaemia, anaemia and autoimmune diseases
- cells also provide a resource for bone marrow replacement therapy in many diseases.
- can now also be transformed into induced stem cells
- Links: Cord Stem Cells
Adult Stem Cells
- Connective Tissue
- Bone marrow - Blood Cells, Osteoclasts, blasts
- Epithelia - Gut, Skin (Epidermis: Immortal Stem Cell)
- Neural?
Epithelium Stem Differentiation
 Epidermis stem cell models - each generation at least 1 "immortal" stem cell - descendants present in patch in future
- Other basal cells - leave basal layer and differentiate
- Committed, born different or may be stem cells
- equivalent to immortal stem cell in character
- mortal in sense that their progeny jostled out of basal layer and shed from 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
Stem Cell Production - Stem Cell Daughter Fates
- Environmental asymmetry
- daughters are initially similar
- 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 two daughters are already have different determinants at time of their birth
- Links: NIH - What are adult stem cells?
Induced Pluripotent Cells
- non-pluripotent cells engineered to become pluripotent (iPSC), a cell with a specialized function ‘reprogrammed’ to an unspecialized state
 embryonic stem cell signalling regulation Yamanaka Factors
A set of 4 transcription factors when introduced into cells induces stem cell formation.[2] These four transcription factors can be expressed from doxycycline (dox)-inducible lentiviral vectors. The only culture difference in iPS cells and human embryonic stem cell culture is that iPS cell culture require 100ng/ml of bFGF in the culture media.
Outline of the MEF reprogramming protocol 1
Outline of the MEF reprogramming protocol 2 | stained with anti-Rex1, Sox2 and SSEA1 antibodies
- OCT4 Transcription factors containing the POU homeodomain
- MYC The MYC protooncogene encodes a DNA-binding factor that can activate and repress transcription. Ectopic expression of c-Myc can also cause tumorigenicity in offspring.
- SOX2 SRY-RELATED HMG-BOX GENE 2
- KLF4 Kruppel-like factor 4, zinc finger protein, transcription factor which acts as both an activator and repressor.
More recently shown that Oct4 together with either Klf4 or c-Myc is sufficient to generate iPS cells from neural stem cells.[3]
Thompson Factor
- Links: Induced Stem Cells | Video - Generating iPS Cells
Stem Cell Markers
In order to carry out research on stem cells, it is important to be able to identify them. A number of different research groups in the late 90's generated several antibodies which specifically identified undifferentiated, differentiating or differentiated stem cells from a number of different sources and species. Note that the nomenclature in some cases is based upon the antibody used to identify the cell surface marker.
Morula cell lines express ES markers[4]
- Every cell surface has specialized proteins (receptors) that can selectively bind or adhere to other “signalling” molecules (ligands)
- Different types of receptors differ in structure and affinity for signalling molecules
- Cells use these receptors and molecules that bind to them as a way of communicating with other cells and to carry out their proper functions in the body
- Stage-specific embryonic antigen (SSEA)-1, -3 and -4 and tumor-rejection antigen (TRA)-1-60 and -1-81, are expressed in specific combinations by undifferentiated pluripotent cells.
- embryonic stem cells, induced pluripotent stem cells, embryonal carcinoma cells, primordial germ cells, mesenchymal progenitors in adult murine bone marrow, and embryonic germ cells.
- Stage-Specific Embryonic Antigen-1 (SSEA-1) cell surface glycan embryonic antigen which has a role in cell adhesion, migration and differentiation and is often differentially expressed during development. Can be identified by Davor Solter monoclonal antibody MC-480 (SSEA-1).
- Stage-Specific Embryonic Antigen-4 (SSEA-4) cell surface embryonic 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. Antigen 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)
- Tumor Rejection Antigen (TRA-1-60) Sialylated Keratan Sulfate Proteoglycan expressed on the surface of human teratocarcinoma stem cells (EC), human embryonic germ cells (EG) and human embryonic stem cells (ES).
- Tumor Rejection Antigen (TRA-1-81) antigen expressed on the surface of human teratocarcinoma stem cells (EC), human embryonic germ cells (EG) and human embryonic stem cells (ES).
- Both TRA antibodies identify a major polypeptide (Mr 240 kDa) and a minor polypeptide (Mr 415 kDa).
- Oct-4 (Pou5f1 – Mouse Genome Informatics) gene has an essential role in control of developmental pluripotency (Oct4 knockout embryo blastocysts die at the time of implantation). Oct4 also has a role in maintaining viability of mammalian germline.
- Stem Cell Antigen 1 (Sca-1) member of the Ly-6 family of GPI-linked surface proteins (Mr 18 kDa) and a major phenotypic marker for mouse hematopoietic progenitor/stem cell subset.
- CD133, AC133, prominin 5 transmembrane glycoprotein (865 aa) expressed on stem cells with hematopoietic and nonhematopoietic differentiation potential.
Expression of Zfp42/Rex1 Gene - used as a marker of undifferentiated stem cells[5]
- regulated by Nanog, Sox2, and Oct4, and by the Wnt pathway
- subject to epigenetic regulation by polycomb complexes and DNA methylation
- Links: PNAS - Molecular markers of ES cells in morula-derived cell lines
Embryonic vs Adult Stem Cells
 Early lineage markers in morulae and blastocysts Embryonic Stem Cell Advantages
- Pluripotency - ability to differentiateinto any cell type.
- Immortal - one cell can supply endless amounts of cells.
- Easily available - human embryos from fertility clinics.
Embryonic Stem Cell Disadvantages
- Unstable - difficult to control differentiation into specific cell type.
- Immunogenic - potential immune rejection when transplanted into patients.
- Teratomas - tumor composed of tissues from 3 embryonic germ layers.
- Ethical Controversy - unethical for those who believes that life begins at conception.
Adult Stem Cell Advantages
- Already ‘specialised’ - induction of differentiation into specific cell types will be easier.
- Plasticity - Recent evidences suggest wider than previously thought ranges of tissue types can be derived.
- No Immune-rejection - if used in autologous transplantations.
- No Teratomas - unlike ES cells.
- No Ethical Controversy - sourced from adult tissues.
Adult Stem Cell Disadvantages
- Minimal quantity - number of isolatable cells may be small.
- Finite life-span - may have limited lifespan in culture.
- Ageing - stem cells from aged individuals may have higher chance of genetic damage due to ageing.
- Immunogenic - potential immune rejection if donor cells are derived from another individual.
Current stem cell research
How to:
- Isolate
- Grow
- Maintain, store
- Differentiate
- Therapeutic uses
References
Textbooks
- The Cell- A Molecular Approach Cooper, Geoffrey M. Sunderland (MA): Sinauer Associates, Inc.; c2000 Stem Cells
Search
Images
Hematopoietic and stromal cell differentiation
stem cell signalling regulation
Epidermis stem cell models
Human embryonic_stem_cell_defined_conditions_01
Human embryonic_stem_cell_defined_conditions_02
Human induced pluripotent_stem_cells
Early lineage markers in morulae and blastocysts
Morula cell lines express ES markers
References
- ↑ <pubmed>20525219</pubmed>| BMC Dev Biol.
- ↑ <pubmed>16904174</pubmed>
- ↑ <pubmed>18594515</pubmed>
- ↑ <pubmed>15917331</pubmed>| PNAS
- ↑ <pubmed>16714766</pubmed>
Journals
Co-ordinator Note
Dr Mark Hill
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ANAT2341 Embryology S2 2011
- --Mark Hill 06:47, 20 July 2011 (EST)
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Course Content 2011
2011 Timetable:
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Embryology Introduction
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Fertilization
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Cell Division/Fertilization
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Week 1 and 2 Development
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Week 3 Development
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Week 1 to 3
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Mesoderm Development
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Ectoderm, Early Neural, Neural Crest
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Trilaminar Embryo to Early Embryo
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Early Vascular Development
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Placenta
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Vascular and Placenta
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Endoderm, Early Gastrointestinal
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Respiratory Development
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Endoderm and Respiratory
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Head Development
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Neural Crest Development
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Head and Neural Crest
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Musculoskeletal Development
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Limb Development
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Musculoskeletal
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Renal Development
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Genital
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Kidney and Genital
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Sensory
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Stem Cells
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Stem Cells
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Endocrine Development
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Endocrine
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Heart
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Integumentary Development
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Heart and Integumentary
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Fetal | Birth and Revision | Fetal
Glossary Links
- Glossary: A | B | C | D | E | F | G | H | I | J | K | L | M | N | O | P | Q | R | S | T | U | V | W | X | Y | Z | Numbers | Symbols | Term Link
Cite this page: Hill, M.A. (2024, April 23) Embryology Lecture - Stem Cells. Retrieved from https://embryology.med.unsw.edu.au/embryology/index.php/Lecture_-_Stem_Cells
- What Links Here?
- © Dr Mark Hill 2024, UNSW Embryology ISBN: 978 0 7334 2609 4 - UNSW CRICOS Provider Code No. 00098G
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