Primary human aortic smooth muscle cells are isolated from black free regions of the human Aorta. It is also isolated in the stain positive from the smooth muscle a-actin. The pulsatile pressure produced by the heart creates a cyclic distention. This particular distinction is happening in the Aorta. This enables the smooth muscle cells to contract again in the aortic wall. The particular changes happening in the arterial wall are completely associated with vascular diseases.
All the vascular diseases, including hypertension, atherosclerosis, etc. They are highly responsible for influencing this process in that region. Therefore, this condition is well capable of studying the role of smooth muscles under disease conditions or normal conditions in vitro. In this article, we will learn about all the essential roles of human aortic smooth muscle cells.
Role and Diseases Associated with Human Aortic Smooth Muscle Cells:
Every patient associated with chronic kidney disease has shown the symptoms of increasing incidence of cardiovascular disease. Due to the high concentration of circulating uremic toxins. It can happen due to the alteration in mineral metabolism and hormone levels. This can be highly responsible for the remodelling of the vascular wall and significant vascular damage. This condition is called medial calcification, an early vascular event in chronic kidney disease patients.
It is completely associated with apoptosis or necrosis. It is also known as transdifferentiation of vascular smooth muscle cells. Human aortic smooth muscle cells Are obtained from bovine and cultured in the presence of increased inorganic phosphate. This is the unique and extensive process used by the researchers for studying these processes. During this process, the human aortic Smooth muscle cells are studied for primary cultures to compare the effects of increased phosphate.
It will help in the treatment method with a serum obtained from uremic patients. Serum induced calcification, phenotypic remodelling and transdifferentiation even with the normal phosphate levels. Even with similar classification kinetics, there are several fundamental differences in osteochondrogenic marker expression. It can also have the fundamental differences in alkaline phosphate induction between phosphate and unique serum treated cells.
Also, we have seen that high phosphate induction has decreased cell viability. On the other hand, uremic serum completely preserved it properly. In other words, the primary culture of this sells with serum from uremic patients has the more informative model. It completely includes vascular calcification secondary to chronic kidney diseases.
Human Aorta Smooth Muscle Cell Treatments and Culture:
Different research laboratories initially isolate human Aorta Smooth Muscle Cells. Then, it proceeds through the purchase from different locations for the differentiation. It is important to understand that it is properly cultured as per the instruction given by the manufacturer. In simple words, all the cells are cultured at 37⁰ C in an atmosphere of 5% Co2. The entire process is done with saturated water. In terms of culture medium, it is done with high glucose Dulbecco’s modified eagles medium supplements.
It is done with the additional 10% fetal bovine serum. It helps to preserve smooth muscle cells into specific growth factors properly. It is also added with penicillin or streptomycin and 0.2% of MycoZap. This entire process will help in preventing mycoplasma contamination. It is important to note that all the culture surfaces are pre coated with a solution containing 1mg/ml fubronectin and 0.2% gelatin. All the mediums are replaced every 48 hours until and unless the cell reaches 50% of the confluence.
This entire process is highly important to maintain the integrity of the cells. After this duration, it is again replaced with daily. The cells are subcultured once they reach 80% to 90% Of confluence. after the sales are confluent, they are carefully transferred to the experimental medium. This medium has the culture medium supplements with 20% of the complete control. In some cases, it also contains uremic human serum. If it is not available, they are also added with DMEM with NaH2PO4, CaCl2 and Na2HPO4.
This will help in reaching the final concentration of 2.5 mM Inorganic phosphate. It also has 2mM Calcium in the medium for the proper integration. This is noteworthy that all the experiments performed using these cells are also in-between passages 3 and 5.
The completely common model of the vitro calcification of Human Aorta Smooth Muscle Cells Culture uses high extracellular phosphate. This helps stimulate hyperphosphatemia, which can be detected in the late cases of chronic kidney diseases. However, in case this model fails to understand the complex metabolic and hormonal alteration. Proper research and findings help in preventing diseases in uremic patients. In other words, several published studies are obtained from bovines.
However, in some cases, we can also find conflicting results due to several species-specific differences in the process. Therefore, it is important to examine the process of phenotypic remodelling and mineralisation closely. This will help in comparison happening in the high extracellular phosphate, which is produced by uremic serum. This serum can be easily obtained from chronic kidney disease patients.