| Background:Myocardial infarction(myocardial infarction, MI) is caused by a blockage of coronary myocardial ischemia and necrosis of a serious hazard to human health disease. Current drug therapy, interventional and surgical treatment of coronary artery bypass graft surgery can only restore regional myocardial blood supply, delaying adverse cardiac remodeling, but not fundamentally solve the myocardium after myocardial infarction to reduce the number of cardiac function of drop this fundamental problem. Our study and others indicate that there is a heart cardiac stem cells(cardiac stem cells, CSCs), which can differentiate into cardiomyocytes, endothelial cells and smooth muscle cells, and can repair damaged heart tissue. Age distribution and gender distribution of this study was to observe the mouse cardiac stem cell subsets from basic research point of view, while each stem cell subpopulations in different parts of the heart in the distribution of the preliminary testing for clinical stem cell therapy of myocardial infarction to find new target.Objective:In this study, from basic research point of view, to observe the mouse heart stem cell subsets age distribution and gender distribution, while each stem cell subpopulations in different parts of the heart in the distribution of the preliminary testing for clinical stem cell therapy of myocardial infarction looking new target.Materials and methods:1. Active use of immunomagnetic sorting C57 mice received cardiac stem cells and the use of flow cytometry to identify cell purity; for sorting the cardiac stem cell lines in cell culture and differentiation ability to observe their orientation.2. Flow cytometry analysis of the content of C57 mice were aged nine cardiac stem cells C57 4-week-old mice and 9-month-old youth; analysis of six-month-old adult female, malemice C57 content of each nine cardiac stem cells; analysis 9 6-month-old healthy adult atrium, left ventricle and right ventricle and ventricular septal cardiac stem cells in C57 mice.Results:1. C57 activity of sorting and identification of cardiac stem cells in mice1.1 C57 mice were isolated and cultured cardiac stem cells successfully1.2 C57 beads were obtained in mice CD45- election law, Sca-1 + stem cells by flow cytometry Purity Test.Lagendorff device perfusion, myocardial enzyme digestion, filtered through 30μm filter small cells were Sca-1 and CD45 antibody after incubation with flow cytometry CD45-, Sca-1 + cells percentage. Been detected, CD45- purified, Sca-1 + cells percentage up to 86.4%, stable cell purification technology.1.3 Isolation of cardiac stem cells have the ability to differentiate moreIsolated from normal adult 6 month old male C57 mouse heart and extracted cardiac stem cells, the culture medium for 14 days and then cells were fixed for immunofluorescence staining, found: stem cells already begin to differentiate a particular cell type. Respectively troponin(troponin I, red), α- smooth muscle actin(α-smooth muscle actin, α-SMA, red), von Willebrand factor(v WF, green) as cardiac cells, fibroblasts and endothelial cells markers(marked in blue with DAPI nuclei). Proof of cardiac stem cells isolated and cultured does have the ability to differentiate into heart tissue cells, namely the separation of cardiac stem cells successfully.2. C57 mouse cardiac stem cells age, sex and location of distribution2.1 C57 mice ckit + cardiac stem cells compare with sca-1 + stem cells in the heart. Select 6-month-old adult male C57 mice 9 were measured using flow cytometry heart ckit + cardiac stem cells and sca-1 + cardiac stem cell content(%). It was found that ckit + CSCs were significantly less than sca-1 + CSCS content.(0.19 ± 0.027 vs 21.37 ± 3.378, P <0.05).2.2 C57 mice of different ages ckit +, sca-1 + cardiac stem cells in comparison.Select 4 weeks and 9-month-old male C57 mice were 9, respectively, as young and old age groups were determined using flow cytometry heart ckit + cardiac stem cells and sca-1 + cardiac stem cell content(%). It was found that the youth group ckit + cardiac cells in the stem(%) more than the older group(0.24 ± 0.036 vs 0.16 ± 0.057, P <0.05); while the oldergroup sca-1 + cardiac cells in the stem(%) was higher than the young group(25.41 ± 4.689 vs 20.41 ± 5.213, P <0.05); as a result of sca-1 + cardiac cells in the stem was significantly higher than ckit + cardiac stem cells, so in general, the older group of cardiac cells in the stem(%) was significantly higher than that of the young group(23.67 ± 6.431 vs 20.21 ± 3.452, P <0.05).2.3 C57 mice of different gender ckit +, sca-1 + stem cells in the heart compared.Select 6-month-old male and a female adult C57 mice 9, respectively, as male and female group, the group, measured using flow cytometry heart ckit + cardiac stem cells and sca-1 + cardiac stem cell content(%). It was found that the female group ckit + cardiac stem cell content(%) with no significant difference between male group(0.19 ± 0.034 vs 0.18 ± 0.045, P>0.05); while the female group sca-1 + cardiac stem cell content(%) and the difference between male group no statistically significant(22.35 ± 3.423 vs 21.90 ± 4.322, P > 0.05); so overall, female and male group content of cardiac stem cells significantly different(21.37 ± 6.231 vs 20.21 ± 2.322, P>0.05).Conclusions:1. MACS method to obtain C57 mice cardiac stem cells have the ability to differentiate into more cardiomyocytes, fibroblasts and endothelial cells of.2. Ckit+ C57 mice were significantly less cardiac stem cells in sca-1+ cardiac stem cells.3. Elderly cardiac stem cells in mice was significantly higher than the youth C57 mice.4. C57 mice of different gender ckit+, sca-1+ stem cells in the heart of no significant difference.5. C57 mice in different parts of the heart ckit+, sca-1+ cardiac stem cells showed no significant difference. |
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