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The International Journal of the Royal Society of Thailand
Volume XI - 2019

Epigenetic is dynamic and reversible process assisting human to adapt to
various environments and diseases. Therefore, epigenetic can be used to
demonstrate pathophysiology of kidney disease in various conditions as well

as a potential targeted therapy development.

Single-cell RNA sequencing

consists of about a million nephrons in and comprised of more than 40
different cell types (Lake et al., 2019; Malone et al., 2018). It is unclear if all nephrons
respond to injury in the same way. Traditional approaches for differentiating cell
types were microscopy and fluorescence-activated cell sorting (FACS). But these
approaches are poor in identification gene expression of single cell. The subsequent
technique was RNA sequencing (RNA-seq) that is able to identify gene expression
by using whole tissue as specimen. The major drawback is that this method can
only identify gene expression from whole cell population, but not from specific cell
types. There were attempts to isolate specific structure of nephrons to analyze the
gene expression of specific cells such as laser capture microdissection technique,
but it still failed to distinguish between glomerular and interstitial cells (Lee et al.,
2015; McMahon et al., 2008).

Figure 4 Workflow of single cell RNA sequencing process. After the tissue was obtained, it was
dissociated into cells. Then, cells were compartmentalized into either plate or microfluidic. RNA
was extracted from cells and convert to cDNA by using reverse transcriptase enzyme. Next, cDNA
was amplified by using polymerase chain reaction (PCR) or in vitro transcription (IVT). Finally,
cDNA was sequenced and analyzed.
Abbreviations: RNA, ribonucleic acid; cDNA, complementary deoxyribonucleic acid

Phatadon Sirivongrangson
Nattachai Srisawat 61

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