Korean J Hematol.  2006 Dec;41(4):250-258. 10.5045/kjh.2006.41.4.250.

Survival and Graft versus Host Disease in Murine MHC Mismatched Hematopoietic Stem Cell Transplantation with Co-injection of Mesenchymal Stem Cells

• Choi S
• Lee DH
• Jeong DC
• Hwang HS
• Chung NG
• Cho B
• Han CW
• Kim HK

• KMID: 2305211
• DOI: http://doi.org/10.5045/kjh.2006.41.4.250

Abstract

BACKGROUND: Intravenous injection of mesenchymal and hematopoietic stem cells (MSCs, HSCs) has the disadvantages of low delivery rate to bone marrow and sequestration of cells in the lung and liver. This study was designed to determine whether there is a relationship between the administration route and dosage of stem cells and GVHD and survival.
METHODS
MSCs were retrieved from five subcultured C3H/10T1/2, cell lines from C3H/He mice. HSCs were transplanted by injecting 1 x 10(7) of bone marrow mononuclear cells and 5 x 10(6) of spleen cells from six to eight week old female C3H/He mice into six week old irradiated female BALB/c mice. The groups were divided into intravenous injection (IV) and intra-marrow (IM) injection groups. IV and IM+MSC groups consisted of mice transplanted with the same bone marrow mononuclear cells and SP, IV and IM groups, with the additional co-injection of 1 x 10(6) MSCs.
RESULTS
Evaluation of all mice, in both groups, showed no difference in GVHD and survival. However, high dose injection with 1 x 10(6) MSCs led to a decreased incidence of GVHD (P<0.05) and improved survival (P<0.01) in both groups.
CONCLUSION
The results of this study showed that the positive effects of MSC on GVHD and survival were primarily dependent on the number of injected cells.

Keyword

Graft versus host disease; Mesenchymal stem cell; Route; Dosage

MeSH Terms

Animals
Bone Marrow
Cell Line
Female
Graft vs Host Disease*
Hematopoietic Stem Cell Transplantation*
Hematopoietic Stem Cells*
Humans
Incidence
Injections, Intravenous
Liver
Lung
Mesenchymal Stromal Cells*
Mice
Spleen
Stem Cells
Transplants*

Figure

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  Fig. 1 The schematic diagram of MHC mismatched murine hematopoietic stem cell transplantation experimental model (MSC, mesenchymal stem cell; IV, intravenous injection; IM, intra-marrow injection).

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  Fig. 2 The severity and pattern of GVHD progression in the experimental group (BALB/c mouse, donor; C3H/He mouse, recipient) using GVHD scoring system of Cooke’s. There is no severe GVHD in all groups and no statistical significance (P＞0.05). (IV group, 1×107 bone marrow mononuclear cells and 3×107 spleen cells intravenously co-injection; IV+MSC group, IV group+1×106 MSC intravenously co-injection; IM group, 1×107 bone marrow mononuclear cells and 3×107 spleen cells intra-marrow co-injection; IM+MSC group, IM group+1×106 MSC intra-marrow co-injection). MSC, mesenchymal stem cell.

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  Fig. 3 Skin and small intestine pathology in BALB/c mouse on 4 days and 8 days after allogenic hematopoietic stem cell transplantation (H&E stain). Skin and small intestine were harvested and prepared for microscopic analysis as described in materials and methods. Interstitial inflammation is observed involving epithelium, mucosae, vessels, parenchyma, and luminal structures on the post transplantation 8 day comparing to the 4th day. Predominantly mononuclear infiltrates and destructions of mucosae and epithelium are also observed.

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  Fig. 4 The severity and pattern of GVHD progression in the intravenous mesenchymal stem cell co-injection hematopoietic stem cell transplantation experimental group. High dose MSC group shows a statistically significant lower GVHD scores from post-hematopoietic stem cell transplantation day 7 onwards compared with the other group (∗P＜0.05). All data were shown mean±SD. (IV+MSC (105) group (n=7), 1×107 bone marrow mononuclear cells+5×10 6 spleen cells+1×105 MSC; IV+MSC (106) group (n=6), 1×107 bone marrow mononuclear cells+5×106 spleen cells+1×106 MSC).

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  Fig. 5 Survival in BALB/c mice in intravenous mesenchymal stem cell co-injection hematopoietic stem cell transplantation experimental group. Whilst all mice in IV+MSC (105) group died in post-transplantation day 8, 4 mice in high dose MSC group survived until post-transplantation day 14, showing improved survival compared with the other group (P＜0.01). (IV+MSC (105) group (n=7), 1×107 bone marrow mononuclear cells+5×106 spleen cells+ 1×105 MSC; IV+MSC (106) group (n=6), 1×107 bone marrow mononuclear cells+5×106 spleen cells+1×106 MSC).

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  Fig. 6 The severity and pattern of GVHD progression in the intra-marrow mesenchymal stem cell co-injection hematopoietic stem cell transplantation experimental group. High dose MSC group shows a statistically significant lower GVHD scores (P＜0.01) from post-hematopoietic stem cell transplantation Day 7 onwards compared with the other group (∗P＜0.01). All data were shown mean±SD. (IM+ MSC (105) group (n=7), 1×107 bone marrow mononuclear cells+5×106 spleen cells+1×105 MSC; IM+MSC (106) group (n=6), 1×107 bone marrow mononuclear cells+5× 106 spleen cells+1×106 MSC).

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  Fig. 7 Survival in BALB/c mice in intra-marrow mesenchymal stem cell co-injection hematopoietic stem cell transplantation experimental group. Whilst all mice in IM group died in post-transplantation day 8, mouse in high dose MSC group survived until post-transplantation day 11, showing improved survival compared with the other group (P＜0.01). (IM+MSC (105) group (n=7), 1×107 bone marrow mononuclear cells+5×106 spleen cells+1×105 MSC; IM+MSC (106) group (n=6), 1×107 bone marrow mononuclear cells+5×106 spleen cells+1×106 MSC).

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