Effects of human umbilical cord mesenchymal stem cells via intramuscular injection on the myocardial ki-67, phh3 and cTnT expression in normal rats

doi:10.3969/j.issn.2095-4344.2014.41.020

Abstract

Abstract:

BACKGROUND: It is unclear whether intramuscular injection of human umbilical cord mesenchymal stem cells will increase regeneration of normal myocardial cells and play adverse effects on normal myocardium.

OBJECTIVE: To observe the effects of intramuscular injection of human umbilical cord mesenchymal stem cells on expression of the myocardial ki-67, phh3 and cTnT in normal Wistar rats.

METHODS: A total of 60 male Wistar rats were divided into six groups randomly: normal group, solution group, supernatant group, low concentration group, moderate concentration group, and high concentration group. These groups were intramuscularly injected different liquid, respectively, as follows: PBS, DMEM, the supernatant of human umbilical cord mesenchymal stem cells, human umbilical cord mesenchymal stem cells with amount of 0.25×10⁵, human umbilical cord mesenchymal stem cells with amount of 1.0×10⁵, human umbilical cord mesenchymal stem cells with amount of 4.0×10⁵. After 4 weeks, each rats received the second same intramuscular injections of human umbilical cord mesenchymal stem cells. Four weeks after the second injection, all rates were killed to obtain myocardial tissues which were fixed, embedded and sectioned. Finally, the ki-67, phh3 and cTnT expressions of the myocardium were detected by immunohistochemical technique.

RESULTS AND CONCLUSION: In the normal group, negative expression of the ki-67 was observed in the caryon of myocardial cells, weak positive expression of the phh3 was observed in the caryon of myocardial cells, and positive expression of the cTnT was observed in the caryon of myocardial cells. Compared with the normal group, the expression of the ki-67, phh3 and cTnT in the myocardium had no significant difference among the

other groups (F＝1.076, 0.167, 0.300; P > 0.05). Human umbilical cord mesenchymal stem cells or the supernatant of human umbilical cord mesenchymal stem cells via intramuscular injection have no effects on the expression of the ki-67, phh3 and cTnT in normal Wistar rats.

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

全文链接：

Key words: umbilical cord, mesenchymal stem cells, injections, intramuscular, myocardium, troponin T, Ki-67 antigen

CLC Number:

R394.2

Wang Qi, Zhang Wen-xiang, Wang Si-ping, Li Zi-pu. Effects of human umbilical cord mesenchymal stem cells via intramuscular injection on the myocardial ki-67, phh3 and cTnT expression in normal rats[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(41): 6671-6677.

Figures/Tables 3

References

[1] Nesselmann C, Kaminski A, Steinhoff G. Cardiac stem cell therapy. Registered trials and a pilot study in patients with dilated cardiomyopathy.Herz. 2011;36(2):121-134.
[2] Forte E, Chimenti I, Barile L,et al. Cardiac cell therapy: the next (re)generation.Stem Cell Rev. 2011;7(4):1018-1030.
[3] Shabbir A, Zisa D, Suzuki G,et al. Heart failure therapy mediated by the trophic activities of bone marrow mesenchymal stem cells: a noninvasive therapeutic regimen. Am J Physiol Heart Circ Physiol. 2009; 296(6): H1888-1897.
[4] Méndez-Ferrer S, Michurina TV, Ferraro F,et al.Mesenchymal and haematopoietic stem cells form a unique bone marrow niche.Nature. 2010;466(7308):829-834.
[5] Tolar J, Le Blanc K, Keating A,et al. Concise review: hitting the right spot with mesenchymal stromal cells.Stem Cells. 2010;28(8):1446-1455.
[6] Gnecchi M, Zhang Z, Ni A, et al.Paracrine mechanisms in adult stem cell signaling and therapy.Circ Res. 2008;103(11): 1204-1219.
[7] Uemura R, Xu M, Ahmad N,et al. Bone marrow stem cells prevent left ventricular remodeling of ischemic heart through paracrine signaling.Circ Res. 2006;98(11):1414- 1421.
[8] Liu Y, Yan X, Sun Z,et al. Flk-1+ adipose-derived mesenchymal stem cells differentiate into skeletal muscle satellite cells and ameliorate muscular dystrophy in mdx mice.Stem Cells Dev. 2007;16(5):695-706.
[9] Shabbir A, Zisa D, Leiker M,et al. Muscular dystrophy therapy by nonautologous mesenchymal stem cells: muscle regeneration without immunosuppression and inflammation. Transplantation. 2009;87(9):1275-1282.
[10] Yang XF. Stem cell transplantation for treating Duchenne muscular dystrophy. Neural Regen Res.2012; 7(22): 1744-1751.
[11] Mueller SM, Glowacki J.Age-related decline in the osteogenic potential of human bone marrow cells cultured in three-dimensional collagen sponges.J Cell Biochem. 2001; 82(4):583-590.
[12] Hsieh JY, Wang HW, Chang SJ,et al. Mesenchymal stem cells from human umbilical cord express preferentially secreted factors related to neuroprotection, neurogenesis, and angiogenesis.PLoS One. 2013;8(8):e72604.
[13] Nartprayut K, U-Pratya Y, Kheolamai P,et al. Cardiomyocyte differentiation of perinatally?derived mesenchymal stem cells. Mol Med Rep. 2013;7(5):1465-1469.
[14] Maureira P, Marie PY, Yu F,et al. Repairing chronic myocardial infarction with autologous mesenchymal stem cells engineered tissue in rat promotes angiogenesis and limits ventricular remodeling.J Biomed Sci. 2012;19:93.
[15] 王思平.肌肉注射异种脐带间充质干细胞对健康Wistar大鼠心肌VEGF、cTnI表达及血清VEGF、HGF、IGF-1和GM-CSF的影响[D].青岛:青岛大学,2012.
[16] 张文祥.Wistar大鼠肌肉注射异种脐带间充质细胞的安全性研究[D].青岛:青岛大学,2012.
[17] Gnecchi M, Danieli P, Cervio E. Mesenchymal stem cell therapy for heart disease. Vascul Pharmacol. 2012;57(1): 48-55.
[18] van den Akker F, Deddens JC, Doevendans PA,et al. Cardiac stem cell therapy to modulate inflammation upon myocardial infarction.Biochim Biophys Acta. 2013;1830(2):2449-2458.
[19] Mohammadi Gorji S, Karimpor Malekshah AA, Hashemi-Soteh MB,et al. Effect of mesenchymal stem cells on Doxorubicin-induced fibrosis.Cell J. 2012;14(2):142-151.
[20] Kajstura J, Leri A, Finato N,et al. Myocyte proliferation in end-stage cardiac failure in humans.Proc Natl Acad Sci U S A. 1998;95(15):8801-8805.
[21] Kajstura J, Leri A, Castaldo C,et al. Myocyte growth in the failing heart.Surg Clin North Am. 2004;84(1):161-177.
[22] 何洪燕,林晓波,应文娟,等.人脐带间充质干细胞经体内定植并向心肌样细胞分化的研究[J].中国输血杂志,2009, 22(3):188-191.
[23] Ozaki K, Yamagami T, Nomura K,et al. Prognostic significance of surgical margin, Ki-67 and cyclin D1 protein expression in grade II canine cutaneous mast cell tumor.J Vet Med Sci. 2007;69(11):1117-1121.
[24] Beltrami AP, Urbanek K, Kajstura J, et al. Evidence that human cardiac myocytes divide after myocardial infarction.N Engl J Med. 2001;344(23):1750-1757.
[25] Ribalta T, McCutcheon IE, Aldape KD,et al. The mitosis-specific antibody anti-phosphohistone-H3 (PHH3) facilitates rapid reliable grading of meningiomas according to WHO 2000 criteria.Am J Surg Pathol. 2004;28(11):1532- 1536.
[26] Tapia C, Kutzner H, Mentzel T,et al.Two mitosis-specific antibodies, MPM-2 and phospho-histone H3 (Ser28), allow rapid and precise determination of mitotic activity.Am J Surg Pathol. 2006;30(1):83-89.
[27] Apple FS, Collinson PO, IFCC Task Force on Clinical Applications of Cardiac Biomarkers. Analytical characteristics of high-sensitivity cardiac troponin assays.Clin Chem. 2012;58(1):54-61.
[28] Katus HA, Remppis A, Scheffold T,et al. Intracellular compartmentation of cardiac troponin T and its release kinetics in patients with reperfused and nonreperfused myocardial infarction.Am J Cardiol. 1991;67(16):1360-1367.
[29] Fishbein MC, Wang T, Matijasevic M,et al. Myocardial tissue troponins T and I. An immunohistochemical study in experimental models of myocardial ischemia.Cardiovasc Pathol. 2003;12(2):65-71.
[30] Heeschen C, Deu A, Langenbrink L,et al. Analytical and diagnostic performance of troponin assays in patients suspicious for acute coronary syndromes.Clin Biochem. 2000;33(5):359-368.
[31] Pagani F, Bonetti G, Panteghini M.Comparative study of cardiac troponin I and T measurements in a routine extra-cardiological clinical setting.J Clin Lab Anal. 2001; 15(4):210-214.