Detecting Technologies And Characteristics Of Telocyte In Mice Heart

Background:Telocyte(TC)is a special type of cell, which is similar to the gastrointestinal Cajal interstitial cell(ICC). It exists in multiple systems or organs in mammals, its characteristics are with long and thin projections(TP) around the cell body. The immunohistochemical staining(IHC)and/or transmission electron microscopy(TEM) are used aboard, but the research about the cardiac TC moves slowly.Objective: To explore the existence and characteristics of TC in the heart of mouse using IHC, primary cells culture and TEM, as contrast group of the gastrointestinal ICC and the bladder TC. The advantages and disadvantages of several detection techniques are compared.Methods: Fifty-nine cases of Kunming mice aged from 10 days to 10 weeks were chosen in this study. ACK2, the antibody of c-kit, was used to identify the myocardial tissue in situ IHC, and the primary cultured cardiac cells. The light microscope, fluorescence microscope and TEM were combined used to observe the express of c-kit. The gastrointestinal ICC and the bladder TC were used as positive control.Results: Operating technique of IHC was without the sterile with the period 2 days. Primary cell culture and immunofluorescence laser confocal microscopy were in the sterile with period of 4-6 days. TEM was without the sterile with the cycle of 15-20 days. No expression of c-kit was found in all of the frozen sections of murine cardiac tissues by IHC. But it positively expressed in the in situ cardiac tissues and the primary cultured cardiac cells by immunofluorescence stain and the TEM. Both the cardiac TC and the ICC or TC of positive control had spindle or irregular bodies, and the mean value of the cell body area of atrium and ventricle TC were 9.18±3.67?m2and 9.17±3.28?m2; while the ICC or TC of small intestine, colon and the bladder were 9.47±5.16?m2,12.12±3.84?m2and 12.30±2.61?m2,and the difference of the area are not significant. There has statistical significance of the TP length between the atrium and ventricle TC(P=0.001),the former is shorter then the latter. There has statistical significance of the TP length among the ICC or TC of small intestine,bladder and atrium TC(P=0.001?0.005),the former two are longer than the latter. There has statistical significance of the TP length between the colon ICC and the ventricle TC(P=0.018),the former is shorter than the latter. TC and ICC in these tissues had 1 to 3 prominent long and thin TP on one single section under the TEM. Cardiac TC with 3 TPs accounted for 44.44 percent, cells with 1 TP 16.67 percent; the small intestine ICC with 3 TPs accounted for 45.45 percent, cells with1 TP 121.68 percent; the bladder TCwith 3 TPs accounted for 46.97 percent, cells with 1 TP 18.19 percent.The TPs between neighboring TC closely linked or wove and intertwined to each other. The diameter of TP is uneven for the crisscrossed expansions and depressions. A kind of long strip dense substance was found between the TP of the ventricle TC and the capillary. The cytoplasm and TP of TC contained a large number of mitochondria, a few lysosome and endoplasmic reticulum.Conclusion: By combined with the method of IHC, primary cell culture and immunofluorescence laser confocal microscopy and TEM, the reliability of cardiac TC detection can improve. TC with the positive expression cell of c-kit was discovered in mice heart. The cell area of TC was similar to the gastrointestinal ICC or the bladder TC. The TP length of ventricle TC is the longest in these TC or ICC, while the atrium TC shortest. It is located in myocardial fibers or around the capillary, TPs of ventricle TC may have some close interrelation with the capillary.