cloning in nature
DESCRIPTION
Cloning in Nature. Parthenogenesis. Monozygotic twins (embryo splitting). (incomplete embryo splitting). Egg Cell and Sperm Cells. Pg. 661. Ca 2+ Wave. Metaphase arrest. Ca 2+ and Egg Activation. SEP. “Calcium wave”. Cleavage Stage Embryos. Pre-compaction. Compaction. - PowerPoint PPT PresentationTRANSCRIPT
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Cloning in Nature
ParthenogenesisMonozygotic twins(embryo splitting) (incomplete embryo
splitting)
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Egg Cell and Sperm Cells
Pg. 661
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Ca 2+ Wave
Metaphase arrest
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Ca2+ and Egg Activation
“Calcium wave”
SEP
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Cleavage Stage Embryos
Pre-compaction Compaction
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Cadherin (Homophilic Cell Adhesion Molecule)
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Blastocycst
Uterus
BlastocystInner Cell Mass (ICM)
50 µmTrophoblast
Blastocoel
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Pre-implantation Development (~ 7 days in humans)
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Mammals Cloned by Embryo Splitting
SheepCattleMousePigHorseRhesus monkey
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Blastomere Isolation
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“Tetra”
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Mammals Cloned by SCNT
SheepMouseCattlePigMule
“Dolly”
“Copy Cat”
HorseGoatRabbitCatDogGaur
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Removal of genetic material from oocyte
Transfer of diploid nucleusfrom adult somatic cell or Embryonic cell into“enucleated” oocyte
Cleavage divisions of“reconstructed” embryo
Oocyte Activation
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Injected somatic nucleus
CondensedChromosomes Pseudo-pronuclei
SCNT in Mice
Ca 2+ injectioncytochalasin
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Removal of genetic material from oocyte
Transfer of diploid nucleusfrom adult somatic cell or Embryonic cell into“enucleated” oocyte
Cleavage divisions of“reconstructed” embryo
Oocyte Activation
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Copy Cat Somatic cell donor“mom”
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Copy Cat Somatic cell donor“mom”
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Inactive X Chromosome (“Barr Body”)
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X Chromosome Inactivation
ICM cells ofBlastocyst
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Copy Cat Somatic cell donor“mom”
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Neural Plasticity
Environment has a significant role in establishing and maintaining neural connections in the brain
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Stem Cell Concept
PluripotentMultipotentUnipotent
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Regenerative Medicine
Stem Cells
Transplant into patients
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Pluripotent Stem Cells
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Mouse Chimeras with Embryonic Stem Cells
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Science 282: 1145-1147November 6, 1998
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Human Embryonic Stem Cell Colony
Stem Cells
Mouse“feeder”cells
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Normal Karyotype in Cultured Human Embryonic Stem Cells
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Telomerase Activity
Cultured Cells Remain Undifferentiated
Undifferentiation “Markers”
Oct4 SSAE4
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Teratoma
Ectoderm Mesoderm Endoderm(gut epithelium)(bone and cartilage)(squamous epithelium)
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Embryonic Stem Cell Signaling Pathways
These Pathways are necessary for maintaining the undifferentiated State
OCT4
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Nature 456: 344-349November 20, 2008
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Science 318: 1917-1920December 21, 2007
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Science 321: 1218-1221August 29, 2008
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iPS Cell Colony from Patient with ALS
Reprogramming Genes: OCT4, SOX2, KLF4, c-MYC
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Neurons from iPS Cells Derived from ALS patient
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Tissues with Adult Stem Cells
Bone marrow/peripheral blood/umbilical cordBlood vesselsBrain/spinal cordSkeletal muscleColonLiverPancreasRetina/corneaSkinDental pulp
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Adult Stem Cell
Multipotent
Life-long
Plasticity?
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Adult Stem Cells
Multipotent
Progenitor Cells
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Stem Cell Niche
Stem Cell
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(Common Lymphoid progenitor)
(Common Myeloid progenitor)
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Signaling Pathways for Self Renewalin Adult Stem Cells
Signals produced with the HSC niche
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Cytkine Signaling via Jak/STAT Pathway
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Cytokines in Hematopoiesis
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Bone Marrow
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(Common Lymphoid progenitor)
(Common Myeloid progenitor)
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Myeloid Leukemia
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Tumor Growth
(~ 100 days/pop doubling)
(~ 7.5 yrs)
(~ 8 yrs)
(~ 10 yrs)
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Myeloid Leukemia
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CD34+
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Myeloid Leukemia
CD34+
CD34+
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Self-renewalProliferation
No Wnt Molecules
(No cell division)
Pathway OFF
INACTIVE
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Self-renewing Proliferation
Wnt Target Genes
Cyclin DNanogSOX2c-mycLIN28BMP4FGFMMP
(family of ~ 20 proteins)
Pathway ON
ACTIVE
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CONSTANT Proliferation
Wnt Target Genes
Cyclin DNanogSOX2c-mycLIN28BMP4FGFMMP
(family of ~ 20 proteins)
Pathway AlwaysON inCancer
Activating Mutations
WntFRATcatenin
Inactivating Mutations
AxinAPC
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Tumor Formation
All Cell Typesin the TumorDifferent Proliferative Potential
Phenotypically Different Cells
Varying Degrees of Differentiation
Different Lifespans
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Implications for Cancer Chemotherapy
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December 4, 2008
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Tumorigenic Potential
~ 0.0001 - 0.1% of Cancer cells are Tumorigenic/Leukemogenic
Assay
Inject tumor cells into SCID miceExamine mice after 8 weeks
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Prolonged Observation periodSeverely immunocompromised micePreincubation of injected tumor cells in ECM proteins
Factors Affecting Outcome of Experiment