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Cells capable of bone production engraft from whole bone marrow transplants in nonablated mice.

Nilsson SK, Dooner MS, Weier HU, Frenkel B, Lian JB, Stein GS, Quesenberry PJ - J. Exp. Med. (1999)

Bottom Line: This was done at the individual cell level in situ, with significant numbers of donor cells being detected by fluorescence in situ hybridization in whole femoral sections.Engrafted cells were functionally active as osteoblasts producing bone before being encapsulated within the bone lacunae and terminally differentiating into osteocytes.Transplanted cells were also detected as flattened bone lining cells on the periosteal bone surface.

View Article: PubMed Central - PubMed

Affiliation: Cancer Center and the Department of Cell Biology, University of Massachusetts Medical Center, Worcester, Massachusetts 01605, USA. s.nilsson@pmci.unimelb.edu.au

ABSTRACT
Allogeneic and autologous marrow transplants are routinely used to correct a wide variety of diseases. In addition, autologous marrow transplants potentially provide opportune means of delivering genes in transfected, engrafting stem cells. However, relatively little is known about the mechanisms of engraftment in transplant recipients, especially in the nonablated setting and with regard to cells not of hemopoietic origin. In particular, this includes stromal cells and progenitors of the osteoblastic lineage. We have demonstrated for the first time that a whole bone marrow transplant contains cells that engraft and become competent osteoblasts capable of producing bone matrix. This was done at the individual cell level in situ, with significant numbers of donor cells being detected by fluorescence in situ hybridization in whole femoral sections. Engrafted cells were functionally active as osteoblasts producing bone before being encapsulated within the bone lacunae and terminally differentiating into osteocytes. Transplanted cells were also detected as flattened bone lining cells on the periosteal bone surface.

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Percentage of male DNA in marrow of individual recipient  female mice determined by Southern blot. Male was taken as 100% and  female as 0%. Loading variability was corrected using an IL-3 probe.
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Figure 2: Percentage of male DNA in marrow of individual recipient female mice determined by Southern blot. Male was taken as 100% and female as 0%. Loading variability was corrected using an IL-3 probe.

Mentions: FISH of femoral marrow sections. Panel A, male control showing positive Y chromosomes (m; deep red/orange). Panel B, female control (f  ); autofluorescence appears yellow. Panel C, transplant recipient 6 wk after transplant of 1.2 × 108 male marrow cells. Sections are viewed using a dual red and green filter, and therefore have no counter-staining. Marrow chimerism was 23.9 ± 2.7% (n = 5), as determined by Southern blot analysis (Fig. 2). Panel D, same as C viewed using a DAPI filter. Note complete lack of auto fluorescence in positive cell. bm, bone marrow; d, positive bone cell of donor origin; e, endosteum; p, periosteum. Bars, 20 μm.


Cells capable of bone production engraft from whole bone marrow transplants in nonablated mice.

Nilsson SK, Dooner MS, Weier HU, Frenkel B, Lian JB, Stein GS, Quesenberry PJ - J. Exp. Med. (1999)

Percentage of male DNA in marrow of individual recipient  female mice determined by Southern blot. Male was taken as 100% and  female as 0%. Loading variability was corrected using an IL-3 probe.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2192923&req=5

Figure 2: Percentage of male DNA in marrow of individual recipient female mice determined by Southern blot. Male was taken as 100% and female as 0%. Loading variability was corrected using an IL-3 probe.
Mentions: FISH of femoral marrow sections. Panel A, male control showing positive Y chromosomes (m; deep red/orange). Panel B, female control (f  ); autofluorescence appears yellow. Panel C, transplant recipient 6 wk after transplant of 1.2 × 108 male marrow cells. Sections are viewed using a dual red and green filter, and therefore have no counter-staining. Marrow chimerism was 23.9 ± 2.7% (n = 5), as determined by Southern blot analysis (Fig. 2). Panel D, same as C viewed using a DAPI filter. Note complete lack of auto fluorescence in positive cell. bm, bone marrow; d, positive bone cell of donor origin; e, endosteum; p, periosteum. Bars, 20 μm.

Bottom Line: This was done at the individual cell level in situ, with significant numbers of donor cells being detected by fluorescence in situ hybridization in whole femoral sections.Engrafted cells were functionally active as osteoblasts producing bone before being encapsulated within the bone lacunae and terminally differentiating into osteocytes.Transplanted cells were also detected as flattened bone lining cells on the periosteal bone surface.

View Article: PubMed Central - PubMed

Affiliation: Cancer Center and the Department of Cell Biology, University of Massachusetts Medical Center, Worcester, Massachusetts 01605, USA. s.nilsson@pmci.unimelb.edu.au

ABSTRACT
Allogeneic and autologous marrow transplants are routinely used to correct a wide variety of diseases. In addition, autologous marrow transplants potentially provide opportune means of delivering genes in transfected, engrafting stem cells. However, relatively little is known about the mechanisms of engraftment in transplant recipients, especially in the nonablated setting and with regard to cells not of hemopoietic origin. In particular, this includes stromal cells and progenitors of the osteoblastic lineage. We have demonstrated for the first time that a whole bone marrow transplant contains cells that engraft and become competent osteoblasts capable of producing bone matrix. This was done at the individual cell level in situ, with significant numbers of donor cells being detected by fluorescence in situ hybridization in whole femoral sections. Engrafted cells were functionally active as osteoblasts producing bone before being encapsulated within the bone lacunae and terminally differentiating into osteocytes. Transplanted cells were also detected as flattened bone lining cells on the periosteal bone surface.

Show MeSH