Is there anyone working on cholesterol depletion by methyl-beta cyclodextrin?
I saw a couple of papers where cells (Jukart T, HEK-293, H1299, H23, A549, and Hela cells) were treated with varying concentrations of MBCD i.e 0mM - 10mM for 30mins in 37degrees. Apparently, some cells are viable at until 50% cholesterol depletion compared to the untreated cells. However, I haven't seen anyone reference where cells were depleted after 30mins time point.
I am working on exosome production and need to keep cells depleted for at least 48 hrs to get enough EV, so could you please point me to any references where cells have been depleted with MBCD for more than 12hrs. My preliminary data shows that HEK-293T be indeed depleted and viable for a longer period and I am not sure not if it's because of the high proliferation of 293T or just the SV-40 antigen T in 293T.
Thanks.
Cholesterol is an essential component of animal cell membranes, playing a pivotal role in modulating membrane fluidity, activity of membrane-bound proteins, and the formation of lipid rafts. Given its importance, cholesterol depletion can have profound effects on cell function and viability. The duration for which cells can survive without cholesterol varies based on the cell type, the extent of cholesterol depletion, and the specific experimental conditions used.
When researchers study cholesterol depletion in cultured cells, they often use agents like methyl-β-cyclodextrin (MβCD) to extract cholesterol from the plasma membrane. Here's what happens to cells upon cholesterol depletion:
As for how long cells can be kept with cholesterol depletion, it's variable:
- Some cells might start showing functional effects within minutes to hours after cholesterol depletion.
- Others might survive for several hours to days but could have compromised functions.
- Some cells, especially those that rely heavily on membrane-associated processes or lipid rafts for signaling, might be more sensitive and exhibit functional impairments or death more quickly.
It's essential to note that complete cholesterol depletion is challenging to achieve, and most experimental setups lead to partial depletion. The extent and duration of depletion are crucial variables determining cell survival.