IMM250H1 Lecture Notes - Cd38, Methyl Cellulose, Ex Vivo
Document Summary
Imm334 developmental immunology fall 2013 first lecture. Increase frequency of src by using limiting dilution assay intrafemoral injection and scid/nod mice. Nod/scid repopulating cells (src) within the cd34+cd38- population. Day 1 mice injected cells grew better in engraftment. Day 4 cells and vector incubated with cytokines, then injected culturing led to fewer. In vitro assay based on progenitors" ability to proliferate and differeniatte into colonies in solid meadia (methylcellulose based) in response to cytokine stimulation, then count colonies. Southern blot detects specific dna molecules out of a mix. Immunodeficient mice show strain-specific differences in the engraftment of human stem cells. Test this using southern blot (pre-pcr times) for alu sequence to detect presence of human cells. Result: alu (and thus human cells) detected only if engraft into nod/scid mice. In scid mice, no b or t cells, normal nk cells.
