Pattern of localization of primitive hematopoietic cells in vivo using a novel mouse model  Rachel Bolante-Cervantes, Shunan Li, Amrik Sahota, Jay A. Tischfield, Ted Zwerdling, Peter J. Stambrook  Experimental Hematology  Volume 27, Issue 8, Pages 1346-1352 (August 1999) DOI: 10.1016/S0301-472X(99)00064-8

Figure 1 Clearance of 14C adenine from APRT wild-type and APRT null mice. Two wild-type and two APRT null mice were injected intraperitoneally with 2 μCi per gram weight of 14C adenine. One hour later and at subsequent intervals urine (A) and blood from orbital sinus bleeding (B) were collected and counted. Open squares and open diamonds represent individual wild-type mice; open triangles and open circles represent APRT null mice Experimental Hematology 1999 27, 1346-1352DOI: (10.1016/S0301-472X(99)00064-8)

Figure 5 Autoradiographic detection of donor cells in tissues of nonirradiated and sublethally irradiated recipients. Nonirradiated or sublethally irradiated APRT null recipient mice were injected intravenously with 106 APRT-positive c-kit–positive enriched pluripotential cells. After 8 weeks, recipient mice were injected intraperitoneally with 14C-adenine and 48 hours later, tissues were fixed, paraffin embedded, sectioned, and coated with emulsion. After developing for 7 days at 4°C in a light-tight container, slides were developed, stained with hematoxylin and eosin, and analyzed by light microscopy. Representative tissue sections are shown at 20× magnification. wt ctrl = nontransplanted, wild-type mice injected with 14C-adenine 48 hours prior to analysis; null ctrl = nontransplanted, APRT null mice injected with 14C-adenine 48 hours prior to analysis; post-transplant = representative sections of tissues from sublethally irradiated mice. Tissues sections from nonirradiated mice are not shown, but demonstrated a similar level of detection sensitivity Experimental Hematology 1999 27, 1346-1352DOI: (10.1016/S0301-472X(99)00064-8)

Figure 2 Isolation of c-kit–positive cells from bone marrow of wild-type 129sv mice. Bone marrow from femurs of APRT wild-type mice was flushed and mononuclear cells were interacted with fluoroisothiocyanate-conjugated control antibody (A) or a c-kit monoclonal antibody (B) and sorted by FACS. The bar indicates the displacement of c-kit–positive cells that were collected and used as donor cells Experimental Hematology 1999 27, 1346-1352DOI: (10.1016/S0301-472X(99)00064-8)

Figure 3 PCR detection of donor cells as a function of time in peripheral blood of recipient mice. At biweekly intervals after introduction of wild-type c-kit–positive donor cells into APRT null recipient mice, the presence of donor cells in peripheral blood was assessed by PCR. Blood was obtained by orbital sinus bleeding. The PCR product is a 635-bp fragment characteristic of the wild-type APRT gene. Lanes A, B, C, and D represent results from individual sublethally irradiated recipients. Lanes wt and null represent results from wild-type and null control mice, respectively Experimental Hematology 1999 27, 1346-1352DOI: (10.1016/S0301-472X(99)00064-8)

Figure 4 PCR detection of donor cells in tissues of recipient mice. Eight weeks after introduction of wild-type c-kit–positive donor cells, APRT null recipient mice were sacrificed. The presence of donor cells in liver, bone marrow, kidney, spleen, lung, and thymus was analyzed by PCR. The PCR product is a 635-bp fragment characteristic of the wild-type APRT gene. Lanes A, B, C, and D represent results from individual sublethally irradiated recipients. Lanes E, F, G, and H represent results from nonirradiated recipients. Lanes wt and null represent results from wild-type and null control mice, respectively Experimental Hematology 1999 27, 1346-1352DOI: (10.1016/S0301-472X(99)00064-8)