Creation of a Human ImmuneSystem in a SCID Mouse ModelUsing Artificial Bone MarrowBy: Andrea CantúBackground InformationStem cells and Tissue EngineeringTissue engineering is the design and growth of cell tissues andorgans outside of the human body.Stem cells are the primitive cells present in all organisms thatcan give rise to more stem cells and are the progenitors of allspecialized cells in the body.Adult stem cells found in infants, umbilical cords, placentas, teeth, and adult tissue can renew themselves and have limited ability to differentiate intoother types of cells, usually only the cell types found in their tissueof origin. Little is known about adult stem cell identities, how theydifferentiate and mature, and their complete level of plasticity.Regenerative tissue engineering has many potential impacts onmedicine and science.SCID MiceBoneMarrowSCID: severe combined immune deficiencymice are unable to make T cells or B cellsrare and spontaneous mutation on chromosome 16.the inner mass of a large bonecontains two types of stem cells: hematopoietic andstromalmatrix used was designed to be like human boneSEM ICC Scaffold Stromal Cells on ICC ScaffoldHuman bone marrowSCID B Cell ModelDissection and ChoppingWash PBSDigestion (Trypsin 0.1%)TriturationFiltration (200m)Cell Isolation from MouseSpleen and Thymus TissueImplanted MatrixMouse ThymuSCID Mouse ModelMethod• Was seeded withCD34+/CFSE• 7 day incubationon back of nudeSCID mouseCD150 (red)HLA class I (red)CD133 (red)CD34 (red)400X 400X400X400X630XCD123 (red)MatrixM1M2M3M1M2M3M1M2M3200XSpleenThymusBone Marrow64%33%91%29%66%CFSEM1M2M1M2M1M2M1M2M1M2M1M2M1M2M1M2M1M2M1M2M1M2M1M2IgD630X400XIgMFrom Implanted Bone Marrow CellsFrom Implanted Cord Blood Cells99%controlcontrol IgD CD19CD10 CD4100%91%99%75%25%93%7%control IgD CD19control CD10 CD499%98%34%66%99%57%43%74%26%Spleen9%CD3From Implanted Bone Marrow CellsFrom Implanted Cord Blood CellsCD19CD3CD8CD4Dissected thymusThymusM1M2M1M2M1M2M1M286%14%89%11%78%22%81%19%CD3CD8 CD19CD4M1M273%27%M1M2M1M2M1M285%15%80%20%94%6%ConclusionThis SCID mouse model created a source of maturecells and immature cells that may continue todifferentiate.Not all of the progenitor cells differentiated andmatured, making this a good source for immune cellproduction.CFSE staining showed migration of cells andexhibited the proliferation of the human cellsthroughout the mice.Many future applications of this informationReferencesH.J. Rippon and A.E. Bishop. Embryonic StemCells. Cell Prolif., Vol 37, 23-34 (2004).University of California Center for AnimalAlternativeshttp://www.vetmed.ucdavis.edu/Animal_Alternativeshttp://www.medterms.comhttp://www.stemcells.nih.govSpecial Thanks to:Dr. Joan Nichols, PhD., Dr. Joaquin Cortiella, MPH. MD. JeanNiles, Sonia Bynum, Ryan Yancy, and Esther Valdivia.Support for thisproject comes fromthe United StatesDepartment ofDefense (DefenseAdvancedResearch
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