Metabolic adaptation and fragility in healthy 3-D in vitro skeletal muscle tissues exposed to [CFS] and Long COVID-19 sera, 2025, Mughal+

[DMissa]

There is a lot of talk in this thread about muscle culture and seahorse etc, I haven't gotten to read the paper yet but useful things to look into would be a) whether activity matched controls b) passaging/handling of the cultures (primary muscle cell phenotypes change with doublings - is that what has been used? havent looked in detail at what was cultured) c) absolute basal oxygen consumption rate values not being negative or close to 0 - if reported as normalised to cell number or stained DNA fluorescence or total protein this might be harder to glean d) not a huge amount of variability after ionophore injection (if used, ia assume so) this can cause cells to swell up and detach which confounds measurements, many aspects of the protocol have to be optimised to avoid it such as instrument mixing protocol, choice or presence of adherence matrix, incubation times, etc

sorry to offload homework, under the pump

my interactions with the lead author leave me confident that pitfalls would have been avoided but yes those are the first things that spring to mind when it comes to evaluating this kind of work. there are others but too technical to explain briefly

 

[DMissa]

I also wonder how this relates to the study by @chillier which found no difference in oxygen consumption rate when ME/CFS serum was added to myoblasts. Do these studies contradict each other?

is it the same same cell type and what are the differences in how they were handled would be my questions (have absolutely no time to look)

 

[jnmaciuch]

We have a student in the lab doing this, I sat down with her and went through it a few days ago, actually. It was pretty objective and automated. That mito morphology package was part of the workflow, i just dont remember the full suite she was using. So it's possible this work here is also objective and automated

Good to know!