Influences of CD133+ cells on human umbilical cord blood mononuclear cell transplantation for treating heart failure

doi:10.3969/j.issn.2095-4344.2016.28.014

Abstract

Abstract:

BACKGROUND: Cell purification can eliminate the biological variability of cells, providing new insight into cell regeneration therapy.
OBJECTIVE: To study the Influence of CD133⁺ cells on human umbilical cord blood mononuclear cell transplantation for treatment of heart failure.
METHODS: Human cord blood mononuclear cells were isolated using lymphocyte separation medium method, and CD133⁺ and CD133^- cells were sorted using immunomagnetic beads at a cell density of 1×10⁸/L. Forty Sprague-Dawley rats were randomized into five groups: sham group, model group, CD133⁺ cell group, CD133^- cell group and mononuclear cell group. Animal models of heart failure were made using intraperitoneal injection of isoproterenol in all the groups except for the sham group. Rats in the CD133⁺ cell group and CD133^- cell group were given 1 mL CD133⁺ cells plus 1 mL PBS and 1 mL CD133^- cells plus 1 mL PBS via the tail vein, respectively. Rats in the mononuclear cell group were given 1 mL CD133⁺ cells plus 1 mL CD133^- cells via the tail vein, and those in the sham and model groups given 2 mL PBS via the tail vein. After 4 weeks, cardiac pathology, degree of myocardial fibrosis and colonization of CD133⁺ cells in myocardial tissues were observed.
RESULTS AND CONCLUSION: Hematoxylin-eosin staining showed that myocardial tissues arranged disorderly in the model group, but regularly in the sham group; myocardial disorders were mildest in the CD133⁺ cell group, successively followed by the mononuclear cell group, and severest in the CD133^- cell group and model group. Masson staining showed that in the model group, collagen fibers were proliferated, arranged irregularly and even broken, while in the sham group, the collagen fibers were less in number and arranged in order. Additionally, there was less reduction in collagen fibers and milder myocardial disorders in the CD133⁺ cell group compared with the other groups. Area of collagen fibers was increased significantly in all the groups except for the sham group (P < 0.05), but this increment was the minimal in the CD133⁺ cell group. Findings from immunohistochemistry and immunofluorescence staining showed that there were no CD133⁺ cells in the myocardial tissues of rats. Therefore, our data indicate that compared with the mononuclear cell transplantation, CD133⁺ cell transplantation exerts superiorities in relieving myocardial damage and reducing myocardial fibrosis. However, CD133⁺ cells are not colonized in the myocardial tissue after transplantation.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Cord Blood Stem Cell Transplantation, Heart Failure, Membrane Glycoproteins, Tissue Engineering

CLC Number:

R394.2

Ma Li, Xie Yi-xu, Chang Yu, Yao Lei . Influences of CD133⁺ cells on human umbilical cord blood mononuclear cell transplantation for treating heart failure[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(28): 4196-4202.

Figures/Tables 2

2.1 实验动物数量分析 40只SD大鼠均进入结果分析，中途无脱落。 2.2 各组大鼠心脏组织苏木精-伊红染色结果模型组大鼠出现心肌排列紊乱，假手术组心肌排列整齐，CD133+细胞组、CD133-细胞组和单个核细胞组的心肌紊乱程度介于模型组和对照组之间。CD133+细胞组、CD133-细胞组和单个核细胞组比较，CD133+细胞组心肌紊乱程度最轻，单个核细胞组其次，CD133-细胞组最重，CD133-细胞组和模型组接近，见图1。 2.3 各组大鼠心脏组织的Masson染色结果 Masson染色后红细胞、纤维素和肌纤维呈红色，弹力纤维呈棕色，胶原纤维呈蓝绿色。镜下观察发现，模型组出现胶原纤维增多，排列紊乱，胶原纤维断裂等现象，假手术组胶原纤维很少，且排列整齐，CD133+细胞组、CD133-细胞组和单个核细胞组的胶原纤维增多和紊乱程度介于模型组和对照组之间。CD133+细胞组、CD133-细胞组和单个核细胞组比较，CD133+细胞组心肌胶原纤维减少，紊乱程度最轻，单个核细胞组其次，CD133-细胞组最重，CD133-细胞组和模型组接近。模型组、CD133+细胞组、CD133-细胞组和单个核细胞组的胶原纤维面积均明显高于假手术组(P < 0.05)，与模型组比较，CD133+细胞组和单个核细胞组胶原纤维面积减小(P < 0.05)，模型组和CD133-细胞组相近，差异无显著性意义(P > 0.05)。CD133+细胞组胶原纤维面积少于单个核细胞组(P < 0.05)，见表1，图2。 2.4 各组大鼠心脏组织的免疫组化染色结果免疫组化染色后，深棕色为特异性染色，蓝色为细胞核，显微镜下观察发现，各组大鼠心肌组织中均未发现特异性染色细胞，考虑脐血CD133细胞没有在心肌组织中定植，见图3。 2.5 各组大鼠心脏组织的免疫荧光染色结果免疫荧光染色后，特异性抗体经FITC标记后，在荧光显微镜下可见到翠绿色荧光，深蓝色为细胞核，显微镜下观察各组大鼠心肌组织中均未发现与细胞核重合的特异性荧光细胞，与免疫组化结果一致，脐血CD133细胞没有在心肌组织中定植，见图4。"

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