Figure 3
Figure 3. BM reconstitution with uterine GFP+ blast: 12-week follow-up in vivo. (A) Schematic representation of the in vivo BM reconstitution. (B) A single, GFP+ CD34+/c-kit− uterine blast colony (mixed with wild-type [C57BL/6] BMCs) was used to reconstitute the BM of irradiated, C57BL/6 mice (N = 15). Representative scatter plots illustrating percentages of GFP+ cells in the BM, blood, or spleen of the chimeric recipients (C; by FACS; N = 5). RT-PCR confirmed GFP mRNA expression in the chimeric BM and spleen (D). Control indicates wild-type BMCs (negative control); and positive, GFP+ transgenic BMCs (positive control). Recipient BMCs were cultured in MethoCult media. GFP+ colony-forming units were observed at day 7 (E). Representative micrographs illustrating coexpression of GFP and hematopoietic markers (CD45, CD4, Mac-3, CD13, CD16, and CD8) in BM isolates from the recipients (F, white arrows). DAPI indicates nuclear stain.

BM reconstitution with uterine GFP+ blast: 12-week follow-up in vivo. (A) Schematic representation of the in vivo BM reconstitution. (B) A single, GFP+ CD34+/c-kit uterine blast colony (mixed with wild-type [C57BL/6] BMCs) was used to reconstitute the BM of irradiated, C57BL/6 mice (N = 15). Representative scatter plots illustrating percentages of GFP+ cells in the BM, blood, or spleen of the chimeric recipients (C; by FACS; N = 5). RT-PCR confirmed GFP mRNA expression in the chimeric BM and spleen (D). Control indicates wild-type BMCs (negative control); and positive, GFP+ transgenic BMCs (positive control). Recipient BMCs were cultured in MethoCult media. GFP+ colony-forming units were observed at day 7 (E). Representative micrographs illustrating coexpression of GFP and hematopoietic markers (CD45, CD4, Mac-3, CD13, CD16, and CD8) in BM isolates from the recipients (F, white arrows). DAPI indicates nuclear stain.

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