4/11/2023 0 Comments Scid mouseMethods of analysis.Ĭlonality of the tumor cells grown in SCID mice was demonstrated by in situ hybridization using an adaptation of published methods. Absorbance at 450 nm was determined on a Auto-Reader II ELISA reader (Ortho Diagnostic Systems, Raritan, NJ). After a final washing, 50 μL/well of OPD solution (DAKO, Carpinteria, CA) containing 3% H 2O 2 was added. Fifty microliters per well of streptavidin-horseradish peroxidase were added to each well after washing and allowed to bind for 1 hour. After washing three times with PBS/Tween, plates were incubated with 50 μL/well of biotinylated Ab (affinity-purified antihuman κ and λ light chains at 0.5 μg/mL and antihuman IgA and IgG at 0.2 μg/mL) for 1 hour. Standards consisting of each purified Ig were added to the appropriate plates at concentrations ranging from 0.4 to 300 ng/mL. Serial dilution of samples in PBS-containing 1% BSA (50 μL/well) were incubated at room temperature for 2 hours. The plates were washed three times in PBS containing 0.5% (vol/vol) of Tween 20 and washed one more time with blocking buffer containing 4% bovine serum albumin (BSA) in PBS. Plates were coated with 50 μL/well of primary antihuman κ and λ (5 μg/mL) and antihuman IgA and IgG (10 μg/mL) and incubated overnight at 4☌. Antibodies were purchased from The Binding Site (San Diego, CA). Levels of human IgG, IgA, κ, and λ light chains were determined by enzyme-linked immunosorbent assay (ELISA). © 1998 by The American Society of Hematology. We conclude that the SCID-hu mouse is a favorable host for studying the biology and therapy of myeloma and that a normal bone marrow environment can support the growth of myeloma cells. Active angiogenesis was apparent in areas of myeloma cell infiltration the new endothelial cells were of human origin. The human microenvironment was infiltrated with Epstein-Barr virus-negative monoclonal myeloma cells of the same clonality as the original myeloma cells. Myeloma-bearing mice had high levels of monotypic human Igs, high blood calcium levels, increased osteoclast activity, and severe resorption of the human bones. Engraftment of myeloma cells was followed by detectable human Ig levels in the murine blood. Myeloma cells from the bone marrow of 80% of patients readily grew in the human environment of SCID-hu mice. The SCID-hu mouse harbors a human microenvironment in the form of human fetal bone. Progress in unraveling the biology of myeloma has suffered from lack of an in vitro or in vivo system for reproducible growth of myeloma cells and development of disease manifestations.
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