The differentiation of renal progenitor cells is a task which is critical for many researchers and medical workers who need to help patients whose kidneys are irreversibly damaged.
The study carried out by Minyong Kang and Yong-Mahn Han (2014) is aimed at showing how human induced pluripotent stem cells (HIPSCs) can be differentiated in nephron progenitor cells (NPCs). This study includes a specific protocol describing the steps implemented by the scholars.
In turn, one should explain these procedures. These results can have significant implications for many medical workers, especially if the efficiency of this method is improved. This is why one should examine this technique in greater detail.
Overall, this process consists of several important steps. In particular, NPCs have to pass the following stages:
- primitive streak;
- intermediate mesoderm;
- nephron progenitors;
- renal tubular cells and glomerural podocytes (Kang & Han, 2014, p. 4).
At first, one should mention that the stems were placed in Dulbecco’s Modified Eagle Medium since this medium is critical for the growth of cells. Apart from that, this medium contains such elements as penicillin-streptomycin, serum replacement, and some non-essential amino acids (Kang & Han, 2014, p.2).
Additionally, it is important to change the first step involved in conventional cardiomyocyte differentiation (Kang & Han, 2014, p.2). In particular, the researchers had to place the cells in the serum-free environment. Additionally, some changes have to be made at the point when it is necessary to derive mesoderm from primitive streak.
At this point, it is necessary to replace bone morphogenetic protein (BMP) 4 with BMP 7 (Kang & Han, 2014, p.2). To some degree, these substitutions are important for improving the growth of these cells. These are some of the main steps that should be taken at the initial stages.
Additionally, nephron progenitors have to be placed on dishes that are coated with fibronectin. Furthermore, the medium has to be changed during every two days. Apart from that, much attention should be paid to tubulogenic assay which is important for the formation of endothelial cells.
Another part of this process is polymerase chain reaction. This step is critical for generating various copies of a specific DNA sequence. In this case, the task of researcher is to determine if the RNA isolated from new cells contains glyceraldehyde-3-phosphate dehydrogenase or GAPDH gene (Kang & Han, 2014, p. 2).
It is the main marker that should be identified. This procedure can show whether the previous differentiation steps were correct.
Furthermore, one should consider the use of immunocytochemistry. This procedure was necessary to determine whether the grown cells include specific markers or proteins that are typical of nephron progenitors. For instance, one can speak about such markers as PAX2 and WT1 (Kang & Han, 2014, p. 2).
The final stage of this process includes the differentiation of glomerural podocytes and tubular cells. At this stage, Minyong Kang and Yong-Mahn Han (2014) rely on the protocols and procedures developed by other researchers.
Additionally, it is necessary to apply such a method as podocytes induction which is important for the generation of SYN-positive cells (Kang & Han, 2014, p 1). These are some of the main processes which were involved in this experiment.
It should be mentioned that this experiment can be implemented successfully provided that at every stage, researchers apply specific solutions that are important for the growth of cells. Furthermore, the combination of these solutions is important for increasing the rate of induction.
As it has been said before, the authors modify some of the solutions that were applied before. This is one of the issues that should be taken into account.
Overall, this process is based on the premise that the differentiation of NPCs is important for the restoration of damaged tissues. It should be mentioned that kidneys include 10 types of different cells; in turn, NPCs are important for the regeneration of these cells.
The proposed method is more cost-effective and less dangerous because it is much easier to obtain induced stem cell which can later be used for the treatment of various patients. This is one of the advantages that can be identified.
Admittedly, the process developed by researchers has certain limitations. At first, one should consider scholars conduct only in-vitro experiments (Kang & Han, 2014). In other words, they examine cells that are placed outside their biological context. Therefore, scientists cannot tell if these cells can function properly within a human body.
Additionally, one should mention that this procedure has limited induction efficiency. At present, it is necessary to improve this procedure in order to make it more applicable for medical purposes.
Nevertheless, despite these limitations, the procedure that has been described should be considered by other medical researchers. In particular, these findings can be rather valuable because they can be applied for the treatment of various diseases. In particular, this procedure can offer an alternative source of cells.
This argument is applicable to those cases when kidney is damaged irreversibly. These are the main points that can be made.
Reference
Kang, M., & Hang, Y. (2014). Differentiation of Human Pluripotent Stem Cells into Nephron Progenitor Cells in a Serum and Feeder Free System. PLOS, 9(4), 1-11.