In many experiments where HK-2 cells were stimulated with TGF-β at 10 ng/mL for 48 hours, I observed that even in the control group, the baseline expression level of fibrosis proteins like FN1 and αSMA was very high, almost comparable to the TGF-β group. The culture medium for HK-2 cells is DMEM/F12 + 10% FBS + 1% PS. I am uncertain if I made any mistake that caused the HK-2 cells to start fibrosis on their own. If anyone could provide some advice, I would greatly appreciate it. Thank you!
The high baseline fibrosis level in HK-2 cells, which are human kidney proximal tubular epithelial cells, can be attributed to several factors:Cell Line Characteristics: HK-2 cells inherently express certain fibrotic markers due to their origin and the nature of the cell line. This can lead to higher baseline levels of fibrosis-related proteins and genes.Culturing Conditions: The conditions under which HK-2 cells are cultured, including the type of medium, serum, and growth factors used, can influence their fibrotic state. For instance, high glucose concentrations, oxidative stress, or certain cytokines present in the culture medium can induce fibrotic responses.Epigenetic Factors: Long-term culture and passaging of HK-2 cells can lead to epigenetic changes that may upregulate fibrotic pathways.Intrinsic Response to Stress: HK-2 cells might have an inherent response to in vitro stress conditions, such as changes in oxygen levels, that can trigger fibrotic mechanisms.
Hello Hanzhi Liang
The cell culture medium formulation and its modification can affect the HK-2 cell behavior during TGF-β1 induced in in vitro EMT. The HK-2 cell line is a proximal tubule cell line, and proximal tubule is the major player in glucose reabsorption. So, HK-2 and other proximal tubule cell lines are used for studying mechanisms of diabetic kidney disease which is the foremost cause of renal failure.
You may want to refer to the article attached below for more information.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10794419/
Your media for routine culture would be DMEM/F12 (high glucose). But for treatment media, you may use DMEM/F12 (low glucose).
The study in the paper attached below suggest that the levels of vimentin and α-SMA increased in the high glucose group compared with the control group. More information may be obtained from the article.
Article High-glucose induced toxicity in HK-2 cells can be alleviate...
Some more articles for your reference.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9956654/
Article Empagliflozin Protects Against Proximal Renal Tubular Cell I...
Article High glucose induces apoptosis via upregulation of Bim expre...
https://www.por-journal.com/journals/pathology-and-oncology-research/articles/10.3389/pore.2021.629829/full
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