PNB 3260 Lecture Notes
PNB 3260 Lecture Notes 3260
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This 3 page Class Notes was uploaded by AnnaCiara on Saturday January 23, 2016. The Class Notes belongs to 3260 at University of Connecticut taught by Dr. Conover in Spring 2016. Since its upload, it has received 57 views. For similar materials see Stem Cell Biology in Physiology at University of Connecticut.
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Date Created: 01/23/16
PNB 3260 Stem Cell Biology Week 1 Notes Lecture 1 January 19, 2016 Stem cells can self renew -->committed cell -->differentiated cells Many diseases involve the inappropriate death of cells Example: Parkinson’s Disease ~1 million new cases a year, very prevalent in U.S. Example: Type I Diabetes (About 10% of diabetes patients), expensive treatment Example: Heart disease affects tens of millions in the U.S. and leads to millions of fatalities Stem cells Helpful for novel drug based therapies Learning about disease Applications of stem cells 1. Replace cells lost to degenerative disease. 2. Patient-specific stem cells can be used to screen for therapies & understand disease etiology. 3. Understand the role of stem cells in cancer progression. Ethical questions Should surplus embryos be used? Is "destroying" embryos to make cell line ethically okay? Controversy about human-animal chimera - effects on the brain of the animal when human brain cells are transplanted Debates and discussions may thwart advances in stem cell research Lecture 2 January 21, 2016 Fundamentals of Developmental and Stem Cell Biology: Patterning in the embryo After fertilization of oocyte there is surrounding zona pellucida it's an extracellular matrix material that doesn't grow in size cell eventually becomes primarily nucleus and less cytoplasm reason for zona pellucida: o restriction of additional sperm entering activated when sperm penetrates o keeps it's own distinct formation compaction stage is when there is junction and connection between cells - not as loose - this stage could be differentiated with a cell membrane marker o important for attachment "hatching" when embryo breaks through zone pellucida then embryo can implant in the embryo wall - don't want embryo to break out before this because it will lead to ectopic pregnancy in fallopian tube Blastocyst cells pressed to one side and there is a fluid filled cavity Trophoblast: the outer cells that will form placenta and umbilical cord Embryoblast: the inner cells (2 kinds) one form endoderm o includes the inner cell mass (ICM) that will form embryo primitive endoderm will be yolk sac remainder will be epiblast cells location/positioning will distinguish future function then forms into gastrula 7 days in humans Gastrula amionic next to embryo ectoderm, medoderm, endoderm, germ cells inserts into uterine wall and expands Hypoblast is the primitive endoderm (aka yolk sac) Stem cell therapy remove trophoblasts and culture them to be self renewing and pluripotent then give them different things to induce differentiation mouse cultures don't work for humans Growth factor conditions promote different stem cell lines from early mouse and human embryos "Passages" means division so many times then you must split the plates FGF and activin needed for ESCs in humans and not mice EpiSCs = embryonic stem cells already primed and set to their eventual fates/differentiation ESCs (embryonic stem cells) not primed primed and naive stem cells inhibit FGF and GSK3 to make naive ESCs o completely undetermined state o want to take EpiSCs to Naive ESCs to be at full capacity Cellular development potential Totipotent - can make the entire organism - includes cells created from the very first divisions after fertilization Pluripotent - able to form complete embryo (all 3 germ layers) Multipotent - able to form various cell types, but usually only from one germ layer Pluripotency Examples: o Morula o inner cell mass o epiblast o primordial germ cells o germline stem cells Somatic cells need the activation of a transcriptional regulatory network (NANOG, SOX2, OCT4, TCF3 and others ) o nuclear transfer o direct reprogramming derivation of pluripotent stem cells won't get pluripotent cells after the epiblast Gastrulation and Germ layers ectoderm, medoderm, endoderm crest (blastopore lip) form directly across from where sperm penetrates time where all the cells are migrating start forming 3 layers ectoderm - skin, PNS and CNS, neural tube and somites forming medoderm - muscle, connective tissue, blood, bone endoderm - gut and respiratory system, relies on mesoderm each layer influences the other layers Germ layers ectoderm --> epidermis and nervous system medoderm --> bones, muscle, connective tissues, blood tissues endoderm --> gut epithelium and respiratory system Functional Criteria for Pluripotent Stem cells in vitro differentiation to all 3 germ lineages or remain as stem cells Active pluripotency transcriptional regulators (demethylated OCT4 and NANOG promoters) teratoma formation: all 3 layers form when injected into another animal o **Used to test pluripotency of human embryonic stem cells Chimera formation with subsequent germline transmission (mouse only) tetraploid complementation assay (mouse only) (only ESC, NT-ESC and iPSC) Active pluipotency transcriptional regulators = demethylated OCT4 and NANOG promoters Pluirpotent ESCs: Chimaera formation yellow color is host generated cells eventually became a whole living mouse completely derived from those pink cells Pluripotent ESCs: tetraploid Complementation placenta can be 4N 4N ones won't survive so inject fESCs IVF has nothing to do with testing of embryonic stem cells Nuclear transfer generate iPS cells using transcription factors Nobel prize in medicine Gurdon's discovery 1962 o using frog o nuclear transfer o transfered an interestinal cell into enucleated egg and it sometimes would form new tadpole o grown in vitro or in vivo?give b to itself.... Yamanka's discovery o screen transcription factors to see what would convert a somatic cell back into a pluripotent cell o factors: KLF4, Sox2 2 more Ethical concerns human-mouse chimeras: are the animals then partically considered human? o Study transplanted human glial cells into mouse and mouse had increased cognitive ablities generate complete human organs in mouse o ethics of using animals as human organ machines
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