Display Settings:

Items per page

Results: 6

1.
Fig. 4.

Fig. 4. From: Defective osteogenesis of the stromal stem cells predisposes CD18-null mice to osteoporosis.

CD18-/- mice exhibit normal osteoclastic activity. (A) In vivo osteoclastic activity was determined based on the serum concentration of C-terminal telopeptides of type 1 collagen. No significant difference in the total in vivo osteoclastic activity was observed between WT and the deficient mice (P = 0.68, n = 3 mice). (B) The number of mature osteoclasts in the femurs of the WT and CD18-/- mice was determined by tartrate-resistant acid phosphate staining. Similar numbers of osteoclasts were observed in WT and CD18-/- mice (P = 0.13, n = 4 mice). (Magnification: ×200.)

Yasuo Miura, et al. Proc Natl Acad Sci U S A. 2005 September 27;102(39):14022-14027.
2.
Fig. 1.

Fig. 1. From: Defective osteogenesis of the stromal stem cells predisposes CD18-null mice to osteoporosis.

CD18 is expressed by the BMSSCs. (A) Dual-color FACS analyses of the P0 murine BMSSCs, using mAbs specific for CD18, CD14, Sca-1, and CD34. (d) Isotype-matched IgGs were used as controls. (b) Lack of contaminating macrophages within P0 BMSSCs is demonstrated by the negative CD14 staining of CD18+ BMSSCs. CD18 is coexpressed with Sca-1 (a) but not CD34 (c). The percentages of each population were shown in the quadrants. (B) Analysis of CD18 expression in murine and human BMSSCs by immunoblot, using a polyclonal Ab specific for CD18 cytoplasmic domain. CD18 expression was decreased in both murine and human BMSSCs upon consecutive cell passages. Equal protein loading was confirmed by reprobing with mAbs specific for β-actin or α-actinin. The data shown are representative of two independent experiments

Yasuo Miura, et al. Proc Natl Acad Sci U S A. 2005 September 27;102(39):14022-14027.
3.
Fig. 3.

Fig. 3. From: Defective osteogenesis of the stromal stem cells predisposes CD18-null mice to osteoporosis.

Phenotypic comparisons between CD18-/- mice and their WT sex-matched littermates. (A) Faxitron analysis demonstrated a decreased bone density in the femurs of 5-week-old CD18-/- mice (KO, Right) as compared with their WT counterparts (Left). (B) Dual x-ray absorptiometry analysis of the femurs from 15-week-old mice showed a statistically significant difference in BMD between WT (left bar) and CD18-/- (right bar) mice (P = 0.025, n = 4). (C) Hematoxylin/eosin staining on the metaphysis area of the femurs showed diminished trabecular bone structure in CD18-/- (Right) mice as compared with WT mice (Left). (Magnification: ×400.) (D) Representative images of the distal femur metaphysis on microcomputed tomography analysis revealed decreased bone volume, trabecular bone number, trabecular bone thickness, and an increased trabecular bone space. (E) Alizarin red and Alcian blue double skeletal staining for bone (red) and cartilage (blue) of 1-week-old mice. KO, CD18-/-

Yasuo Miura, et al. Proc Natl Acad Sci U S A. 2005 September 27;102(39):14022-14027.
4.
Fig. 5.

Fig. 5. From: Defective osteogenesis of the stromal stem cells predisposes CD18-null mice to osteoporosis.

CD18 deficiency compromises BMSSC differentiation but not proliferation. (A) In vitro mineralization induction of BMSSCs. Mineralization of the CD18-/- BMSSCs, determined by Alizarin red S staining (magnification: ×10), was significantly lower than that of WT cells (P = 0.00003, n = 3 mice). The total mineralized area by WT cells was assigned to 100%. (B) Cell adhesion. BMSSCs were allowed to adhere for 3 h at 37°C. After washing, the adherent cells were counted manually based on 10 randomly picked view fields and expressed as a percentage of WT cell adhesion. The CD18-/- BMSSCs adhered poorly compared with the WT cells (P = 0.03, n = 4). (C) Proliferation of BMSSCs was evaluated by BrdUrd incorporation, and the percentage of BrdUrd+ cells was determined manually by counting 10 representative fields. CD18-/- BMSSCs proliferate better than the WT controls (P = 0.01, n = 3 mice). (D) CFU-F assay. The number of BMSSC colonies obtained from 106 bone marrow cells was significantly increased for the CD18-/- mice (P = 0.0064, n = 6 mice). (Magnification: ×10.)

Yasuo Miura, et al. Proc Natl Acad Sci U S A. 2005 September 27;102(39):14022-14027.
5.
Fig. 6.

Fig. 6. From: Defective osteogenesis of the stromal stem cells predisposes CD18-null mice to osteoporosis.

Defective osteogenic capability of the CD18-/- BMSSCs. (A) Cbfa1 expression and Smad2 phosphorylation by immunoblot. Compared with WT cells, the CD18-/- BMSSCs exhibited decreased Cbfa1 expression (Upper) and enhanced response toward TGF-β stimulation, as indicated by Smad2 phosphorylation (Lower). Equal protein loading was verified by reprobing with an α-actinin-specific mAb. The data shown are representative of three independent experiments. (B) Retroviral mediated expression of recombinant CD18. The CD18-/- BMSSCs were infected with retroviral supernatants encoding either full-length CD18 or the cytoplasmic tail-truncated CD18 (CT-) for 6 days, and expression of recombinant CD18 was determined by immunoblot, using an anti-CD18 cytoplasmic tail Ab, which does not react well with CD18 (CT-). Noninfected WT and CD18-/- BMSSCs were included as controls, and protein loading was shown by reprobing for α-actinin. (C) Bone formation in vivo. WT, CD18-/-, as well as the retroviral-infected CD18-/- BMSSCs with either CD18 or CD18 (CT-) were mixed with hydroxyapatite/tricalcium phosphate and then implanted s.c. in nude mice. The BMSSC-mediated bone formation was analyzed 7 weeks after implantation by hematoxylin/eosin staining. B, newly formed bone; HA, hydroxyapatite/tricalcium phosphate; CT, connective tissues. (Magnification: ×200.) (D) Restoration of the osteogenic capability of the CD18-/- BMSSCs. The amount of bone formation in C was quantified by using the software nih image (http://rsb.info.nih.gov/nih-image) based on five representative areas and was expressed as a percentage of bone formation by WT BMSSCs. Bone formation by CD18-/- BMSSCs was significantly decreased compared with the WT BMSSCs (P = 0.008, n = 4 mice). The defective osteogenesis of CD18-/- BMSSCs was rescued by expression of CD18 (P = 0.0001, n = 4 mice) but not CD18 (CT-) (P = 0.39, n = 4 mice)

Yasuo Miura, et al. Proc Natl Acad Sci U S A. 2005 September 27;102(39):14022-14027.
6.
Fig. 2.

Fig. 2. From: Defective osteogenesis of the stromal stem cells predisposes CD18-null mice to osteoporosis.

Multipotent differentiation capability of the STRO-1bright/CD18+ BMSSCs. (A and B) Enrichment of the STRO-1bright/CD18+ BMSSC population by cell sorting. Fresh human bone marrow mononuclear cells were subjected to cell sorting using both STRO-1 and CD18 as specific markers. Two distinct cell populations (R2, STRO-1bright/CD18+; and R3, STRO-1bright/CD18-) were collected (A) and subjected to CFU-F assays (B). The STRO-1bright/CD18+ cells formed much higher number of colonies than the STRO-1bright/CD18- cells. (C) Phenotypic characterization of the sorted human STRO-1bright/CD18+ BMSSCs was performed by FACS analysis using different lineage-specific mAbs (filled in red). Their corresponding isotype-matched nonimmune IgGs were used as controls (bold line). The numbers given indicate the percentages of the positive cell population. Lack of macrophage contamination of the STRO-1bright/CD18+ BMSSCs was demonstrated by the negative staining of either CD14 or CD45. (D) Differentiation potential of the STRO-1bright/CD18+ BMSSCs. The sorted STRO-1bright/CD18+ cells were cultured in vitro under adipogenic induction condition for 2 weeks (a), osteogenic induction condition for 3 weeks (b), and chondrogenic induction condition for 3 weeks (c). Oil Red O staining demonstrates the generation of lipid-laden adipocytes (yellow arrows) (a); Alizarin Red S staining shows mineral deposits (blue arrows) made by osteogenic cells (b); and Alcian blue staining of the cartilage matrix deposition (green arrows) demonstrates the chondrogenic differentiation in aggregate cultures (c). The data shown are representative of two independent experiments. (Magnification: a and b, ×400; c, ×64.)

Yasuo Miura, et al. Proc Natl Acad Sci U S A. 2005 September 27;102(39):14022-14027.

Display Settings:

Items per page

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk