Stem Cell Engineering Flashcards
Characteristics of SCNT
- NUCLEUS taken from Somatic cell of pt and injected into OOCYTE of a DONOR replacing the oocyte nucleus
- Blastocyst generated from this hybrid oocyte
- ES cells are isolated
Biochemical Comparisons of ES cell character with other types suggest what?
Candidate genes
What can be added to cultured ESC to make a NEURON
Retinoic Acid only
What are the Key determinants of ES cell Character?
Gene Regulatory Proteins
What can be added to cultured ESC to make a SMOOTH Muscle Cell?
- DiButyryl cAMP
+
- Retinoic Acid
Problems with SCNT
- Inefficient
(May need 100+ eggs) - Technically damaging
- Not available in many hospital
Purpose of SCNT
- Combines cloning and ESC technology to produce cells that are COSTOM for the patient
What do ES cells give rise to when Injected into Host animal?
- Bone
- Cartilage
- Skin
- Respiratory lining
- Gut
- Nerves
“Bill Cosby Seems Really Guilty Now”
Challenges of regenerative medicine
- Proper integration into tissue/organ
- Tissue rejection
- Genetic/Epigenetic Aberrations
What can be added to cultured ESC to make an ASTROCYTE or OLIGODENDROCYTE?
- Fibroblast growth factor
(FGF) - FGF-2 and Epidermal Growth Factor (EGF)
- FGF-2 and Platelet-derived Growth Factor
What stage of the embryo development should one harvest for Pluripotent stem cells?
Blastocyst
- Use the ICM to plate and culture to make the Pluripotent stem cells
What does NOT reform from skin of burn patients?
- Hair follicles
- Sebaceous glands
Downside of ES-Like cells
- Yield LOW
- NOT Identical to ES cells
What can be added to cultured ESC to make a MACROPHAGE?
- Macrophage Colony Stimulating factor
2. IL-1 and IL-3
Components of the artificial matrix
- Collagen and GAG
- Lined with thin membrane of SILICONE rubber on outside
What migrates towards the artificial skin and reform the CT
- Fibroblast
- Blood Capillaries
Use of SC to Repair the SKIN
- Cultured Epidermal Cells from healthy skin regions of patients used to isolate stem cells
- Used to repopulate the damaged areas
Why are ES Cells good for artificially making new tissues?
- Pluripotency well established
- Good growth properties in vitro
- ES cells have ability to differentiate into a wide range of tissues if provided the right set of conditions
SCNT used to treat Diabetes
- Pluripotent cell turned into PANCREATIC Stem cell
- Injected into Pancreas
- Stem -> Multipotent -> Pancreatic Cell
What developmental capacity of stem cell differentiation is best suited for reverse engineering?
Pluripotent Cell
- Embryonic Stem Cells
Alternative Approaches: To induce ES Cell-Like Properties
Induced Pluripotent Stem cells (iPS)
Immediate replacement of skin repair
- Use from Cadaver
- Artificial matrix
For preventing water loss
What are the 4 regulatory proteins that make the somatic cells into ES-Like cells?
- Sox2
- Oct3/4
- Klf4
- Myc
Stem Cell Treatment for Leukemia
- Haematopoietic SC taken and multiplied in culture
- Transplanted into patient
- Inside
- Pluripotent -> Multipotent -> Myeloid Progenitor cells -> RBC
