Yonsei Med J.  2013 Jan;54(1):238-245. 10.3349/ymj.2013.54.1.238.

Effect of Human Parathyroid Hormone on Hematopoietic Progenitor Cells in NOD/SCID Mice Co-Transplanted with Human Cord Blood Mononuclear Cells and Mesenchymal Stem Cells

• Lim YJ
• Hwang K
• Kim M
• Cho YH
• Lee JH
• Lee YH
• Seo JJ

• KMID: 1776945
• DOI: http://doi.org/10.3349/ymj.2013.54.1.238

Abstract

PURPOSE
We evaluated the effect of human parathyroid hormone (hPTH) on the engraftment and/or in vivo expansion of hematopoietic stem cells in an umbilical cord blood (UCB)-xenotransplantation model. In addition, we assessed its effect on the expression of cell adhesion molecules.
MATERIALS AND METHODS
Female NOD/SCID mice received sublethal total body irradiation with a single dose of 250 cGy. Eighteen to 24 hours after irradiation, 1x107 human UCB-derived mononuclear cells (MNCs) and 5x106 human UCB-derived mesenchymal stem cells (MSCs) were infused via the tail vein. Mice were randomly divided into three groups: Group 1 mice received MNCs only, Group 2 received MNCs only and were then treated with hPTH, Group 3 mice received MNCs and MSCs, and were treated with hPTH.
RESULTS
Engraftment was achieved in all the mice. Bone marrow cellularity was approximately 20% in Group 1, but 70-80% in the hPTH treated groups. Transplantation of MNCs together with MSCs had no additional effect on bone marrow cellularity. However, the proportion of human CD13 and CD33 myeloid progenitor cells was higher in Group 3, while the proportion of human CD34 did not differ significantly between the three groups. The proportion of CXCR4 cells in Group 3 was larger than in Groups 1 and 2 but without statistical significance.
CONCLUSION
We have demonstrated a positive effect of hPTH on stem cell proliferation and a possible synergistic effect of MSCs and hPTH on the proportion of human hematopoietic progenitor cells, in a xenotransplantation model. Clinical trials of the use of hPTH after stem cell transplantation should be considered.

Keyword

Umbilical cord blood; parathyroid hormone; bone marrow niches

MeSH Terms

Animals
Bone Marrow/metabolism
Cell Proliferation
Female
Fetal Blood/*cytology
Flow Cytometry
Hematopoietic Stem Cell Transplantation
Hematopoietic Stem Cells/*drug effects
Humans
Leukocytes, Mononuclear/*cytology
Mesenchymal Stem Cell Transplantation
Mesenchymal Stromal Cells/*cytology
Mice
Mice, Inbred NOD
Mice, SCID
Parathyroid Hormone/*therapeutic use
Stem Cells/cytology
Transplantation, Heterologous
Parathyroid Hormone

Figure

• 

  Fig. 1 Schema of the xenotransplantation experiment. TBI, total body irradiation; CB, cord blood; MNC, mononuclear cell; MSC, mesenchymal stem cell; hPTH, human parathyroid hormone; CBT, cord blood transplantation; CBC, complete blood count; RV, right ventricle; BM, bone marrow; IP, intraperitoneal; NOD/SCID.

• 

  Fig. 2 Mean complete blood cell counts of mice at the fourth, sixth, and seventh week after transplantation. There are no statistically significant differences between the 3 groups. RBC, red blood cell; WBC, white blood cell.

• 

  Fig. 3 Sections of the bone marrow of tibiae taken from mice of Groups 1, 2 and 3 in the eighth week after transplantation (H&E, ×200). The bone marrow cellularity was decreased to approximately 20% in group 1 (A). Bone marrow cellularity is approximately 70 to 80% in Groups 2 (B) and 3 (C). Bone marrow cellularity and trabecular bone formation are greater in Groups 2 and 3 than Group 1.

• 

  Fig. 4 The representative flow cytometric images of 3 groups. (A) Group 1, (B) Group 2, (C) Group 3. MNC, mononuclear cell; MSC, mesenchymal stem cell; hPTH, human parathyroid hormone.

• 

  Fig. 5 Flow cytometric analysis of engrafted human marrow mononuclear cells in bone marrow aspirates. (A) The percentage of myeloid lineage cells was larger in Group 3 than in Groups 1 and 2. (B) The percentage of lymphoid lineage cells was also higher in Group 3, but the difference was not statistically significant.

• 

  Fig. 6 Flow cytometric analysis of human cell adhesion molecules in bone marrow mononuclear cells. There were no statistically significant differences between the three groups.

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