Generation of reconstituted mice using retrovirus vector.

(A) Diagrams of bicistronic retroviral expression vector constructs. (B) Flow cytometric analysis of c-kit⁺ cells 48 hours after retroviral transduction with retrovirus encoding GFP, SDF-1, or intrakine vector. GFP⁺ cells were collected by a cell sorter and subjected to transplantation. (C) Kinetic study on GFP-expressing cells in peripheral blood (■), bone marrow (⋄), spleen (○), and thymus (▵) of each reconstituted mouse transplanted with 1 × 10⁵ GFP⁺ cells per mouse. Each point represents mean percentage ± SE from 4 separate experiments. (D). The mRNA expression level of transferred genes in the reconstituted mice. Total RNA was extracted from splenocytes. Total RNA was reverse transcribed with random primer and amplified using the specific primers for GFP, CXCR4, SDF-1, intrakine, and G3PDH. These results represent 3 independent experiments. ψ Indicates the packaging symbol.