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my title is not very specific. So i will proceed to clarify it. I am trying to make sure that the only blood cells in a sample are Erytocytes, since i want to evaluate their metabolism, I am aware that they don't have Mithocondria and thus catabolize Glucose into Lactate, (I belive I am not making false statements, however if any corrections pop up please give me a heads up.) Thus a way I thought of getting rid of leucocytes was through centrifugation, erythrocytes would be at the bottom of the flask and if We got rid of Supernadant (plama and fine layer of leucocytes), we would have a much purer sample of erythrocytes.

My question is: Do they stil carry out their metabolism, so i can a posteriori evaluate it, even after centrifugation and gettting rid of the plasma. Or evaluating the lactate production afterwards would be impossible? I know there are more easier and methodic ways of getting rid of leucocytes(irradiation that destroys all nucleated cells and leucoreduction by filtracion, for example), but this is somewhat hypothetical, and i need to know the final answer in these circumstances.

Thanks in advance :)

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  • $\begingroup$ @user1136 I apologize for my absolute statement, nothing is ever black or white, I was aware of the HMP shunt, but i failed to see the affect of it in the metabolism since it take 6 molecules of R5P to produce 5 molecules G6P, the net lactate would be bigger if there was no shunt, hypothetically. As for most of the references given about erytrocyte metabolism it was most fun reading them and I discovered things that will almost certainly help me in future studies, even if not, they were very fun to read. $\endgroup$
    – RockishWood
    Mar 27 '20 at 11:36
  • $\begingroup$ @user1136 However i failed to understand one of your previous statements."And 'reserves' of 2,3-DPG can give rise to a 'burst' of ATP production, especially in stored RBC", do you mean that the Lubering-Raport pathway will lower net ATP count, or that a considerable ammount of stored 2,3BPG will enter the Glicolytic pathway in media res are produce more ATP and lactate out of the blue? $\endgroup$
    – RockishWood
    Mar 27 '20 at 11:41
  • $\begingroup$ @user1136 Thanks for the +1! The only implication of the of storage of blood I withdrew from your references was “A practical application of the DPG story was the realization that the increased oxygen affinity of blood during storage is due to destruction of DPG." in the citation classic article by Benesch & Benesch. Much obliged about the information you helped me in on another question I also had! :) $\endgroup$
    – RockishWood
    Mar 27 '20 at 12:29
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Erythrocytes need more than just glucose in order to function and, they also need a extracelullar medium who shares the same osmolarity, so placing your red blood cells into a medium that solely has glucose will be problematic. If you solely want to evaluate the lactic fermentation carried out by erythrocytes on a short period of time, then you can add the blood plasm on your centrifuge tube (which will be in the superior layer) to the newly separated erythrocyte fraction. This way, we make sure your extracelullar medium not only has every metabolite needed, but is also isotonic with respect to your erythrocytes. If you're planning on evaluating this pathway for a considerable amount of time (days, perhaps), then you, most likely, will have to prepare a solution with glucose that is isotonic with respect to your red blood cells.

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  • $\begingroup$ Welcome to Biology.SE! This looks like a good answer, but answers are much more likely to receive a favorable response if you include supporting references (primary literature is best). Without that support, your answer is indistinguishable from opinion. This is a good example of how to format references. ——— You may also want to take the tour and then consult the help pages for additional advice on How to Answer effectively on this site. Thank you! 😊 $\endgroup$
    – tyersome
    Mar 30 '20 at 23:46
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They need glucose, they absorb that from dissolved particles in water (the plasma). So if you isolate the cells, you'd have to put them in a medium with sugar and so on. It would be analogous to the medium they exist in before you centrifuge them, I think.

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  • $\begingroup$ Thanks your answer was was most insightful! $\endgroup$
    – RockishWood
    Mar 26 '20 at 23:01
  • $\begingroup$ If you're interested in what drives osmosis (really, not the "concentration gradient of water"), feel free to have a look here, citeseerx.ist.psu.edu/viewdoc/… $\endgroup$
    – Leif
    Mar 28 '20 at 23:56
  • $\begingroup$ Wow i did not know that, I will certainly look into it! Much Obliged!! $\endgroup$
    – RockishWood
    Mar 30 '20 at 7:04
  • $\begingroup$ I discovered it just a few weeks ago, and it was published not too long ago, in 2009. Have not seen any earlier references. The short story is, the membrane is what drives osmosis, acting like a pump. That might sound weird, it does initially, but water organizes into a 4th state at membrane surfaces (or broadly speaking "contact zones"). This 4th state is analogous to ice but without the hydrogen ions to bond the lattice sheets vertically, those ions are instead excluded (ionized). Colloids affect the thickness of these gel-layers, and a concentration gradient forms for H+. $\endgroup$
    – Leif
    Mar 30 '20 at 7:25

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