1. Cloning in Nature Parthenogenesis Monozygotic twins (embryo splitting)
(incomplete embryo
splitting)

2. Egg Cell and Sperm Cells
Pg. 661

3. Ca 2+ Wave
Metaphase arrest

4. Ca2+ and Egg Activation
SEP
“Calcium wave”

5. Cleavage Stage Embryos
Pre-compaction
Compaction

6. Cadherin (Homophilic Cell Adhesion Molecule)

8. Blastocycst Uterus Blastocyst Inner Cell Mass (ICM) Blastocoel
Trophoblast
50 µm

9. Pre-implantation Development
(~ 7 days in humans)

10. Mammals Cloned by Embryo Splitting
Sheep
Cattle
Mouse
Pig
Horse
Rhesus monkey

11. Blastomere Isolation

12. “Tetra”

13. Mammals Cloned by SCNT Sheep Horse Mouse Goat Cattle Rabbit Pig Cat
Mule
Horse
Goat
Rabbit
Cat
Dog
Gaur
“Dolly”
“Copy Cat”

14. Removal of genetic material
from oocyte
Transfer of diploid nucleus
from adult somatic cell or
Embryonic cell into
“enucleated” oocyte
Oocyte Activation
Cleavage divisions of
“reconstructed” embryo

15. SCNT in Mice Injected somatic nucleus Condensed Chromosomes
Pseudo-pronuclei
Ca 2+ injection
cytochalasin

16. Removal of genetic material
from oocyte
Transfer of diploid nucleus
from adult somatic cell or
Embryonic cell into
“enucleated” oocyte
Oocyte Activation
Cleavage divisions of
“reconstructed” embryo

18. Copy Cat
Somatic cell donor
“mom”

19. Copy Cat
Somatic cell donor
“mom”

20. Inactive X Chromosome
(“Barr Body”)

21. X Chromosome Inactivation
ICM cells of
Blastocyst

22. Copy Cat
Somatic cell donor
“mom”

23. Neural Plasticity Environment has a significant role
in establishing and maintaining
neural connections in the brain

24. Stem Cell Concept
Pluripotent
Multipotent
Unipotent

25. Regenerative Medicine
Stem Cells
Transplant into patients

26. Pluripotent Stem Cells

27. Mouse Chimeras with Embryonic Stem Cells

28. Science 282:
November 6, 1998

30. Human Embryonic Stem Cell Colony
Stem Cells
Mouse
“feeder”
cells

31. Normal Karyotype in Cultured Human Embryonic Stem Cells

32. Cultured Cells Remain Undifferentiated
Telomerase Activity
Undifferentiation “Markers”
Oct4
SSAE4

33. Teratoma Ectoderm Mesoderm Endoderm (squamous epithelium)
(gut epithelium)
(bone and cartilage)

34. Embryonic Stem Cell Signaling Pathways
OCT4
These Pathways are necessary for maintaining the undifferentiated State

35. Nature 456:
November 20, 2008

36. Science 318:
December 21, 2007

37. Science 321:
August 29, 2008

38. iPS Cell Colony from Patient with ALS
Reprogramming Genes: OCT4, SOX2, KLF4, c-MYC

39. Neurons from iPS Cells Derived from ALS patient

40. Tissues with Adult Stem Cells
Bone marrow/peripheral blood/umbilical cord
Blood vessels
Brain/spinal cord
Skeletal muscle
Colon
Liver
Pancreas
Retina/cornea
Skin
Dental pulp

41. Adult Stem Cell
Life-long
Multipotent
Plasticity?

42. Adult Stem Cells
Multipotent
Progenitor Cells

43. Stem Cell Niche
Stem Cell

44. (Common Myeloid progenitor)
(Common Lymphoid progenitor)

45. Signaling Pathways for Self Renewal
in Adult Stem Cells
Signals produced with the HSC niche

46. Cytkine Signaling via Jak/STAT Pathway

47. Cytokines in Hematopoiesis

48. Bone Marrow

49. (Common Myeloid progenitor)
(Common Lymphoid progenitor)

51. Myeloid Leukemia

52. Tumor Growth (~ 10 yrs) (~ 8 yrs) (~ 7.5 yrs)
(~ 100 days/pop doubling)

54. Myeloid Leukemia

55. CD34+

56. Myeloid Leukemia
CD34+
CD34+

57. Pathway OFF No Wnt Molecules INACTIVE (No cell division) Self-renewal
Proliferation
(No cell division)

58. Pathway ON Cyclin D Nanog SOX2 c-myc LIN28 BMP4 FGF MMP
(family of ~ 20 proteins)
Pathway ON
ACTIVE
Wnt Target Genes
Cyclin D
Nanog
SOX2
c-myc
LIN28
BMP4
FGF
MMP
Self-renewing
Proliferation

59. Pathway Always ON in Cancer Cyclin D Nanog SOX2 c-myc LIN28 BMP4 FGF
(family of ~ 20 proteins)
Pathway
Always
ON in
Cancer
Activating Mutations
Wnt
FRAT
catenin
Wnt Target Genes
Cyclin D
Nanog
SOX2
c-myc
LIN28
BMP4
FGF
MMP
Inactivating Mutations
Axin
APC
CONSTANT
Proliferation

60. Tumor Formation All Cell Types in the Tumor
Phenotypically Different Cells
Varying Degrees of Differentiation
All Cell Types
in the Tumor
Different Proliferative Potential
Different Lifespans

61. Implications for Cancer Chemotherapy

62. December 4, 2008

63. are Tumorigenic/Leukemogenic
Tumorigenic Potential
Assay
Inject tumor cells into SCID mice
Examine mice after 8 weeks
~ % of Cancer cells
are Tumorigenic/Leukemogenic

64. Factors Affecting Outcome of Experiment
Prolonged Observation period
Severely immunocompromised mice
Preincubation of injected tumor cells in ECM proteins