J Korean Med Sci.  2007 Apr;22(2):242-247. 10.3346/jkms.2007.22.2.242.

Cotransplantation of Cord Blood Hematopoietic Stem Cells and Culture-Expanded and GM-CSF-/SCF-Transfected Mesenchymal Stem Cells in SCID Mice

  • 1Department of Laboratory Medicine, Dong-A University College of Medicine, 1 3-ga, Dongdaesin-dong, Seo-gu, Busan, Korea. jyhan@dau.ac.kr
  • 2Department of Microbiology, Dong-A University College of Medicine, Busan, Korea.
  • 3Department of Pharmacology, Dong-A University College of Medicine, Busan, Korea.
  • 4Department of Internal Medicine, Dong-A University College of Medicine, Busan, Korea.
  • 5Department of Pediatrics, Hanyang University College of Medicine, Seoul, Korea.


Mesenchymal stem cells (MSC) are multipotent in nature and believed to facilitate the engraftment of hematopoietic stem cells (HSC) when transplanted simultaneously in animal studies and even in human trials. In this study, we transfected culture-expanded MSC with granulocyte macrophage-colony stimulating factor (GMCSF) and stem cell factor (SCF) cytokine genes and then cotransplanted with mononuclear cells (MNC) to further promote HSC engraftment. MNC were harvested from cord blood and seeded in long-term culture for ex vivo MSC expansion. A total of 1 x 10(7) MNC plus MSC/microliter were introduced to the tail vein of nonobese diabetic/severe combined immunodeficiency mice. After 6-8 weeks later, homing and engraftment of human cells were determined by flow cytometry and fluorescence in situ hybridization studies. The total nucleated cell count and the engraftment of CD45+/CD34+ cells and XX or XY positive human cells were significantly increased in cotransplanted mice and even higher with the cytokine gene-transfected MSC (GM-CSF>SCF, p<0.05) than in transplantation of MNC alone. These results suggest that MSC transfected with hematopoietic growth factor genes are capable of enhancing the hematopoietic engraftment. Delivering genes involved in homing and cell adhesions, CXCR4 or VLA, would further increase the efficiency of stem cell transplantation in the future.


Mesenchymal Stem Cell; Hematopoietic Stem Cell Transplantation; GM-CSF; Stem Cell Factor; Engraftment, Transfection; Cord Blood

MeSH Terms

Stem Cell Factor/genetics/*metabolism
Mice, SCID
Mesenchymal Stem Cells/*metabolism
Mesenchymal Stem Cell Transplantation/*methods
Hematopoietic Stem Cell Transplantation/*methods
Granulocyte Macrophage Colony-Stimulating Factors, Recombinant/*metabolism
Graft Survival/*immunology
Genetic Enhancement/methods


  • Fig. 1 Inverted phase contrast microscopic findings of cultured cord blood MSC at passage 2, day 14 (A) (×150). Wright-Giemsa stained smear of the harvested MSC (B) (×1,000).

  • Fig. 2 Representative immunophenotypic analysis of MSC ex vivo cultures from the cord blood. CD45-/CD73+ (95.7%), CD45-/CD 105+ (97.4%), and CD45-/CD166+ cells (94.9%) were evident by flow cytometry.

  • Fig. 3 GM-CSF transfection studies. Microscopic images of phase contrast (A) and fluorescence microscopes (B) before transfection. Micrographs of phase contrast (C) and fluorescence microscopes (D) post transfection day 5. The panel D shows GFP fluorescence, indicating GFP-expressing MSC after transfection (×200).

  • Fig. 4 SCF transfection studies. Microscopic images of phase contrast (A) and fluorescence microscopes (B) before transfection. Micrographs of phase contrast (C) and fluorescence microscopes (D) post transfection day 7. The panel D shows GFP fluorescence, indicating GFP-expressing MSC after transfection (×200).

  • Fig. 5 XY human cells identified from the peripheral blood of NOD/SCID mice 7 weeks after cotransplantation of GM-CSF-transfected cord blood MSC and MNC (×1,000). One orange signal indicates one copy of the X chromosome, and one green signal indicates one copy of the Y chromosome.

  • Fig. 6 Representative CD34+/CD45+ cells (8.7%) detected from venous blood of NOD/SCID mice 7 weeks after cotransplantation of GM-CSF-transfected cord blood MNC and HSC.


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