Murine Bone Marrow Transplant Assay
Objective: To assess the effects of MPLW515L expression on hematopoiesis and myeloproliferative disorder development in mice using a bone marrow transplant model
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Protocol Steps
Bone marrow cell preparation
Isolate and prepare bone marrow cells expressing either MPLW515L or wild-type MPL for transplantation
Note: Cells must be genetically modified to express the target MPL variant
View evidence from paper
“In a murine bone marrow transplant assay, expression of MPLW515L, but not wild-type MPL, resulted in a fully penetrant myeloproliferative disorder”
Bone marrow transplantation
Transplant prepared bone marrow cells into recipient mice
Note: Recipient mice conditioning regimen not specified
View evidence from paper
“In a murine bone marrow transplant assay, expression of MPLW515L, but not wild-type MPL, resulted in a fully penetrant myeloproliferative disorder”
Hematologic assessment
Measure platelet counts in transplanted mice to assess myeloproliferative disorder development
Note: MPLW515L expression resulted in marked thrombocytosis with platelet counts of 1.9-4.0 × 10^12/L
View evidence from paper
“marked thrombocytosis (Plt count 1.9–4.0 × 10 12 /L)”
Spleen assessment
Evaluate spleen size and histology to assess extramedullary hematopoiesis
Note: MPLW515L expression resulted in marked splenomegaly
View evidence from paper
“marked splenomegaly due to extramedullary hematopoiesis”
Bone marrow fibrosis assessment
Evaluate bone marrow histology for reticulin fibrosis
Note: MPLW515L expression resulted in increased reticulin fibrosis
View evidence from paper
“increased reticulin fibrosis”