Korean Circ J.  2008 Sep;38(9):446-454. 10.4070/kcj.2008.38.9.446.

Therapeutic Potential of Umbilical Cord Blood-Derived Mesenchymal Stem Cells in Ischemic Myocardium

• Kim YS
• Ahn Y
• Hong MH
• Park HJ
• Kwon JS
• Lee HJ
• Kim SH
• Jang SJ
• Song CH
• Kim KH
• Hong YJ
• Kim JH
• Park HW
• Jeong MH
• Cho JG
• Park JC

• KMID: 1776344
• DOI: http://doi.org/10.4070/kcj.2008.38.9.446

Abstract

BACKGROUND AND OBJECTIVES
We designed this study to determine the therapeutic potentials of umbilical cord blood (UCB)-mesenchymal stem cells (MSCs), as compared with bone marrow (BM)-MSCs. MATERIALS AND METHODS: MSCs were isolated from UCB and BM. For the in vivo study, myocardial infarction was induced by ligation of the left anterior descending coronary artery (LAD) in rats for 30 min, and this was followed by release; the MSCs were then injected into a designated point around the infarcted area. Echocardiographs were performed two weeks after surgery. For the in vitro study, a cDNA microarray and cytokine array were performed to compare the MSCs from UCB and from BM. Cell migration was assessed by a wound scratch assay, and the level of cardiac ankyrin repeat protein (CARP) was determined by reverse transcriptase-polymer chain reaction (RT-PCR) or Western blot analysis. RESULTS: For the echocardiograph findings, the fractional shortening (FS) was 43.9% in the UCB-MSCs group and it was 38.6% in the BM-MSC group. The ejection fraction (EF) was 79.8% in the UCB-MSC group and it was 72.4% in the BM-MSC group (control FS: 26.2% and the control EF: 56.6%). CARP was one of the highly expressed genes in the UCB-MSCs on the cDNA microarray. The mRNA and the expressed level of CARP protein in the UCB-MSCs were higher than those in the BM-MSCs. The cell migration of the CARP small interfering ribonucleic acid (siRNA) transfected UCB-MSCs was delayed compared to that of the normal UCB-MSCs (p<0.05) CONCLUSION: Our study directly compared the two types of MSCs from UCB and BM, and we suggest that the CARP molecule might be responsible for the motility of UCB-MSCs.

Keyword

Mesenchymal stem cell; Bone marrow; Umbilical cord blood; Myocardial infarction

MeSH Terms

Animals
Ankyrin Repeat
Blotting, Western
Bone Marrow
Carps
Cell Movement
Coronary Vessels
Fetal Blood
Infarction
Ligation
Mesenchymal Stromal Cells
Myocardial Infarction
Myocardium
Oligonucleotide Array Sequence Analysis
Rats
RNA
RNA, Messenger
Stem Cells
Umbilical Cord
RNA
RNA, Messenger

Figure

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  Fig. 1 Mesenchymal stem cells (MSCs) characterization. FACS analysis and a differentiation study were performed on the umbilical cord blood (UCB)-MSCs (A) and the bone marrow (BM)-MSCs (B), and both groups of MSCs expressed the typical MSC markers. Both groups of MSCs (a) were differentiated to osteoblasts (b), chondrocytes (c) and adipocytes (d). FACS: Fluorescence-Activated Cell Sorter.

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  Fig. 2 Echocardiographic parameters. The ejection fraction (EF) and fractional shortening (FS) of the left ventricle in the cardiac ischemia-reperfusion (IR) injured rats that were treated with mesenchymal stem cells. n=15 in each group, *p<0.05 vs. the IR control rats. LVEF: left ventricular ejection fraction, LVFS: left ventricular fractional shortening, UCB: umbilical cord blood, MSC: mesenchymal stem cell, BM: bone marrow.

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  Fig. 3 DAPI-stained mesenchymal stem cells (MSCs) at 14 days. Sections from the engrafted hearts were stained with antibodies against α-sarcomeric actin (α-SA), von Wilebrand factor (vWF), CD31 and connexin43 (Cn×43). Immunofluorescence staining was done to examine if the injected MSCs were differentiated. The sections were stained for α-SA, CD31, vWF and Cn×43. The MSCs were stained with DAPI before injection to detect the region of the myocardium where they homed to. UCB: umbilical cord blood, MSC: mesenchymal stem cell, BM: bone marrow, DAPI: 4'-6-diamidino-2-phenylindole.

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  Fig. 4 cDNA microarray profiles of the umbilical cord blood (UCB)-mesenchymal stem cells (MSCs) and the bone marrow (BM)-MSCs. The pie charts show the percentages of the cDNAs that were expressed in the UCB-MSCs (A) and BM-MSCs (B). cDNA: complementary deoxyribonucleic acid.

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  Fig. 5 Expression of CARP and the cell migration assay of the umbilical cord blood (UCB)-mesenchymal stem cells (MSCs). The higher expression level of CARP in the UCB-MSCs was confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis (A). To knock down the endogenous CARP in UCB-MSCs, 100 nM of siRNA was transfected and the reduced CARP protein level was shown on Western blotting (B). Cell migration was assayed by the wound scratch assay. UCB-MSCs were transfected with scrambled siRNA or CARP siRNA to examine the effect of CARP on cell motility (C). CARP: cardiac ankyrin repeat protein, siRNA: small interfering ribonucleic acid.

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  Fig. 6 Cytokine array of the umbilical cord blood (UCB)-mesenchymal stem cells (MSCs) and bone marrow (BM)-MSCs. The different cytokines between the two groups of MSCs are marked on the blots. IGFBP: insulin-like growth factor binding protein.

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