Creation of a Human Immune System in a SCID Mouse Model Using Artificial Bone Marrow By Andrea Cant Background Information Stem cells and Tissue Engineering Tissue engineering is the design and growth of cell tissues and organs outside of the human body Stem cells are the primitive cells present in all organisms that can give rise to more stem cells and are the progenitors of all specialized 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 into other types of cells usually only the cell types found in their tissue of origin Little is known about adult stem cell identities how they differentiate and mature and their complete level of plasticity Regenerative tissue engineering has many potential impacts on medicine and science SCID Mice SCID severe combined immune deficiency mice are unable to make T cells or B cells rare and spontaneous mutation on chromosome 16 Bone Marrow the inner mass of a large bone contains two types of stem cells hematopoietic and stromal matrix used was designed to be like human bone Human bone marrow SEM ICC Scaffold Stromal Cells on ICC Scaffold SCID B Cell Model Method SCID Mouse Model Implanted Matrix Cell Isolation from Mouse Spleen and Thymus Tissue Dissection and Chopping Wash PBS Digestion Trypsin 0 1 Trituration Filtration 200 m Mouse Thymu Matrix 400X Was seeded with CD34 CFSE 7 day incubation on back of nude SCID mouse CD123 red HLA class I red CD150 red 630X 400X 400X CD34 red 400X CD133 red CFSE Thymus M2 M3 M1 64 33 Splee n 200X Bone Marrow M2 M2 M3 M1 91 M1 29 M3 66 From Implanted Bone Marrow Cells control Spleen IgD CD19 M1 630X M1 99 91 9 control M1 M2 75 100 CD4 25 M1 M2 93 7 M1 M2 From Implanted Cord Blood Cells control 400X IgD CD19 M2 M1 M2 M1 99 34 control IgM M1 CD10 99 IgD M2 M2 M2 66 M1 99 CD10 M1 98 M2 M1 M2 57 CD4 43 M2 M1 74 M2 26 From Implanted Bone Marrow Cells Thymus CD8 M2 78 Dissecte d thymus CD3 M2 M1 M1 CD19 22 14 86 M2 M1 89 11 19 M1 CD4 81 M2 From Implanted Cord Blood Cells CD8 M1 80 CD3 CD3 20 M2 CD4 6 M1 15 85 M1 94 CD19 M2 M1 M2 73 27 M2 Conclusion This SCID mouse model created a source of mature cells and immature cells that may continue to differentiate Not all of the progenitor cells differentiated and matured making this a good source for immune cell production CFSE staining showed migration of cells and exhibited the proliferation of the human cells throughout the mice Many future applications of this information References H J Rippon and A E Bishop Embryonic Stem Cells Cell Prolif Vol 37 23 34 2004 University of California Center for Animal Alternatives http www vetmed ucdavis edu Animal Alternatives http www medterms com http www stemcells nih gov Special Thanks to Dr Joan Nichols PhD Dr Joaquin Cortiella MPH MD Jean Niles Sonia Bynum Ryan Yancy and Esther Valdivia Support for this project comes from the United States Department of Defense Defense Advanced Research Projects Agency
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