J Korean Med Sci.  2006 Dec;21(6):1000-1004. 10.3346/jkms.2006.21.6.1000.

Cotransplanted Bone Marrow Derived Mesenchymal Stem Cells (MSC) Enhanced Engraftment of Hematopoietic Stem Cells in a MSC-dose Dependent Manner in NOD/SCID Mice

• Kim DH
• Yoo KH
• Yim YS
• Choi J
• Lee SH
• Jung HL
• Sung KW
• Yang SE
• Oh WI
• Yang YS
• Kim SH
• Choi SY
• Koo HH

• KMID: 1713108
• DOI: http://doi.org/10.3346/jkms.2006.21.6.1000

Abstract

Transplantation of marrow-derived mesenchymal stem cells (MSCs), expanded by culture in addition to whole bone marrow, has been shown to enhance engraftment of human hematopoietic stem cells (HSCs). Our hypothesis was that there might be an optimum ratio range that could enhance engraftment. We examined the percent donor chimerism according to the ratio of HSCs to MSCs in non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice. We tested a series of ratios of co-transplanted CD34+-selected bone marrow cells, and marrow-derived MSCs into sublethally irradiated NOD/SCID mice. In all experiments, 1 x 10(5) bone marrow derived human CD34+ cells were administered to each mouse and human MSCs from different donors were infused concomitantly. We repeated the procedure three times and evaluated engraftment with flow cytometry four weeks after each transplantation. Serial ratios of HSCs to MSCs were 1:0, 1:1, 1:2 and 1:4, in the first experiment, 1:0, 1:1, 1:2, 1:4 and 1:8 in the second and 1:0, 1:1, 1:4, 1:8 and 1:16 in the third. Cotransplantation of HSCs and MSCs enhanced engraftment as the dose of MSCs increased. Our results suggest that the optimal ratio of HSCs and MSCs for cotransplantation might be in the range of 1:8-1:16; whereas, an excessive dose of MSCs might decrease engraftment efficiency.

Keyword

Hematopoietic Stem Cells; Mesenchymal Stem Cells; Transplantation; Mice, SCID; Engraftment

MeSH Terms

Middle Aged
Mice, SCID
Mice, Inbred NOD
Mice
Mesenchymal Stem Cells/*cytology
Mesenchymal Stem Cell Transplantation/*methods
Humans
Hematopoietic Stem Cells/*cytology
Hematopoietic Stem Cell Transplantation/*methods
Graft Survival/*physiology
Cells, Cultured
Cell Count
Animals
Adult

Figure

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  Fig. 1 Isolation of HSCs from bone marrow and results of FACS analyses. HSCs were isolated by MACS sort system from bone marrow. The purity of HSCs infused to NOD/SCID mice was more than 90%. (A) Isotypes were stained with FITC-anti-mouse IgG1 and PE-anti-mouse IgG1. (B) Dot plots show FITC-anti-human CD34 on x-axis and PE-anti-human CD38 on y-axis. (C) Isolated CD34+ cells were stained PE-anti-human CD34 only on y-axis.

• 

  Fig. 2 Results of MSCs phenotypes by FACS analysis. MSCs that were used in this test were positive for CD29 (BD-Pharmingen, Palo Alto, CA, U.S.A.), CD44, CD90, CD105, CD166, SH2, SH3, and SH4, while negative for CD14, CD34, CD45, CD51-61, CD106 and HLA-DR. Stro-1 was weakly expressed. Blank peaks indicate the isotype of each cell and closed peaks represent expression of each marker.

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  Fig. 3 MSCs enhanced human cell engraftment in bone marrow of NOD/SCID mice in a dose-dependent manner at 4 weeks posttransplant. (A) CD34+ only group (1.0×105 cells). (B) The ratio of CD34+ to MSCs was 1:1 (1.0×105 cells each). (C) The ratio of CD34+ to MSCs was 1:2 (1.0×105:2.0×105 cells). (D) The ratio of CD34+ to MSC was 1:4 (1.0×105:4.0×105 cells). (E) The ratio of CD34+ to MSC was 1:8 (1.0×105:8.0×105 cells). This is representative of 2 analyses in the experiment 2.

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  Fig. 4 Results of contransplantation. In all experiments, the engraftment in bone marrow was increased as the MSCs dose was increased. In experiment 3, the engraftment in the group of a 1:16 ratio is inferior to that of a 1:8 ratio. *Indicates significant statistical difference between the CD34+ only group and a cotransplanted group.

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