ABSTRACT
Objective: To establish a method for isolation, culture and identification of primary human pericardial interstitial cells (PICs). Methods: Human pericardial tissues were cut into 1 mm×1 mm×1 mm sized pieces, digested with type II collagenase, hyaluronic acid enzyme and trypsin to isolate the interstitial cells. The cells were cultured and cell morphology was observed. The cell doubling time, flow cytometry and immunohistochemical staining were applied for cell identilication. Results: Adherent cells were observed 24 h after culturing. The cells reached 80% confluences and could be subcultured after about 10 days. The cells isolated and cultured were fibroblast-like or spindle-like, with obvious nuclti, clear nucleolus, and a greater proportion of nucleus and cytoplasm. Flow cytometry results showed no cell surface markers CD34 and CD45. Immunocytochemistry showed negative CK staining, and positive vimentin and a{script}SMA staining. Cells isolated from 8 of the 10 pericardial tissues were successfully cultured. Conclusion: The present method can effectively isolate and culture primary human PICs, which lays a foundation for further in vitro study of constrictive pericarditis.
ABSTRACT
To establish a neonatal rat model of thoracic aortic dissection by treating pregnant rats with semicarbazide. Methods Implantable capsule osmotic pump with different concentrations of semicarbazide (5, 10, 15, 20, 25, 30, 35, 40 mg/[kg • d])or normal saline were implanted into the abdomen of rats on day 14 of pregnancy. The thoracic aortas of 21 day fetus and neonatal rats were collected. H-E staining was applied to examine the pathological changes under microscope, and the formation of thoracic aortic dissection was analyzed. Results Various degrees of thoracic aortic dissections were observed in neonatal rats after the mother rats were given different concentrations of semicarbazide. At a lowest semicarbazide release rate of 25 mg/kg per day, all the neonatal rats survived and had a high incidence of thoracic aortic dissection. Conclusion A neonatal rat model of thoracic aortic dissection has been successfully established, paving a way for further researches.
ABSTRACT
Objective:To establish a human lung squamous carcinoma cell line and to study its biological characteristics. Methods:Lung squamous carcinoma specimens were freshly resected during operation;the tissues were incubated in vitro and the cell line was named CHLH-1.The biological characteristics of the cells were studied by light microscopy,electron microsco- py,chromosome analysis and transplantation experiment.Results:Cells from the specimens of the primary tumor,the CHLC- 1 cell line and the cells from transplanted tumor possessed the characteristics of malignant squamous epithelium under light and electron microscope.The cell growth curve,doubling time and mitotic index were also observed in vitro.Nuclear chromosome analysis revealed that the tumor was a subtriploid with a mode of 60-68 per cell.Tumor nodes were observed under the skin of nude mice by heterogenic transplantation.Conclusion:The characteristics of the established cell line suggest that it is a newly established human squamous carcinoma cell line.