How to make Platelet Rich Plasma (PRP) with no Kit!

[@satiashaw1857]

Wow! So informative, thank you!!

[@drbnar1]

Thanks a lot Dr.Buford for the technique , one question , how much ACDA is added to each 60 ml of blood drawn initially ?

[@donbufordmd]

right now we are adding 6cc of ACD-A to a 60cc syringe. The final total is therefore 54cc of blood and 6cc of ACD-A

[@drbnar1]

@ thanks for the reply Dr.Buford , wish you all the best

[@texasorthobiologics]

Platelet count (blood)= 214 ; Platelet count(PRP)= 2827. WBC(blood) =3.1 WBC (PRP) = 11.5 GRA(blood)=1.8 GRA(PRP)=1.4 LYM(blood)=1.00 LYM(PRP)= 8.50 MON (blood)=0.3 Mon (PRP) =1.6

[@fuganetti]

So 13.2x concentration factor and a platelet dose of 16.9 billion total in the 6ml? Impressive!

[@tonypatz9387]

Do you add ACDA tin the drawing syringe? If so how much.

[@marce005]

Any reply?

[@shawnisabellalin8021]

Do you need to activate with calcium?

[@donbufordmd]

You can check PRP in a hematology machine. You should first check the platelet range on your machine to make sure it covers platelet count to up to around 3500 or 4000 so you don’t have to dilute the PRP

[@mine213]

Hi, doc, great informative video, do you have a few recs for hematology machines?

[@Dr-789]

Can we check platelets and other components in PRP by hematology machine that uses usually for CBC etc or it can damage hematology machine ? Thanx

[@texasorthobiologics]

yes you can and it will not damage the machine. For higher platelet concentrations you should check the range on your hematology machine to make sure it can handle platelet counts over 1000....and higher in the PRP ranges.

[@xgcdarkninja911]

Why acd after the first centrifuge?

[@donbufordmd]

to prevent clotting

[@20573867291823047859]

It seems to me it would simplify the process and collect more platelets by using a gel separator tube. After a single spin, you could aspirate the top 80-90% of the plasma. Then you could re-suspend the remaining plasma and platelets in the first tube (thanks to the separator). This should get nearly all of the platelets since you dont have to eyeball it, stopping at the RBC.

[@texasorthobiologics]

Tried...gel separator tubes typically have a very low platelet recovery percentage ...about 50% worse than the technique shown here. There are many reasons for this...but the easiest way to prove it is to run blood at the same time on the two techniques and get the data... we have done that.

[@marcinkowalewski4000]

You are correct. Gel will allow easy collection of platelet-enriched plasma (2 x the baseline max). My father makes PRP in syringes (on daily basis). He actually prepares liquid PRF with 4 to 8 x baseline platelet count (and verifies this with the analyzer). He says, it is the best platelet extract and most economical. We created a syringe clipper so he can clip the flanges automatically and put syringes to a common centrifuge.

[@donbufordmd]

@@marcinkowalewski4000I have tested the gel tubes….they do not provide the flexibility of any two spin system. Additionally is well documented by multiple unbiased doctors that the gel systems have lower Platelet Recovery percentages than the single or two spin techniques without gel separator tubes. Tropocells is the only test tube system that returned a reasonable platelet dose from a 60cc blood draw

[@donbufordmd]

@@marcinkowalewski40002x baseline is not even considered PRP in a clinical setting. Ie. Going from a platelet count of 250 to 500 still means the platelet dose is not therapeutic for virtually any application regardless of how many injections you give. Looked at the RESToRE trial or the PEAK trial. Both online. 2021 and 2022 publication dates I believe

[@shawnisabellalin8021]

You had acda in the blood draw, and you add acda again for the second spin?

[@donbufordmd]

Not every kit protocol does that....but yes, I do add 1.5cc of ACD-A into the test tube for the second spin to make sure there is no platelet aggregation. We have tested this and done it this way for 7 years or so.

[@pedromanuelalvarez7693]

How much ACDA do you add to the initial 60 cc blood draw?

[@wreamdeaver8931]

How long can the blood sit before spinning for plasma?

[@texasorthobiologics]

if you mean after drawing the blood....3-4 hours if handled properly

[@donbufordmd]

Activation is not ecesssry for PRP to work. PRP is activated pretty rapidly upon. Injection anyway

[@zigzag8162]

Is this LR or LP prp?

[@texasorthobiologics]

It can be either. If you take none of the "red" it is LP. If you take about 1cc of the "red" from each 30cc test tube that would make it LR.....taking more red makes it more "LR" and will also raise the hematocrit in proportion.

[@zigzag8162]

@@texasorthobiologics Ok, no RBCs going into the second spin. But, how much of the buffy coat to you include going into the second spin?

[@Dr-789]

Good but u left a lot in first tube which is first PRP ...so my advise to you as dont discord too much in tube after first spin...

[@donbufordmd]

Unfortunately that is simply not true when we actually test that supposition. After the first spin the red is nearly 100% RBC When making LR-PRP we don’t want any of that. After many thousands of doses and a few published papers on this and BMC, I think we have a pretty reliable way to do this. Thanks for taking the time to comment

[@Dr-789]

@@donbufordmd U did not understand me...i am not saying ,not to do double spin...i am actually saying dont left a lot of plasma in tube after first spine as i seen the video...bcoz still there are a lot of platelets in bottom of plasma layer even after first spine which will yield better PRP after second spine....so dont discord plasma after first spin at bottom and take that precious plasma that is close to RBC layer ...thanx

[@donbufordmd]

@@Dr-789 I understand, thanks for the clarification. The typical platelet recovery percentage with the technique shown is 80% which is good......When the goal is truly leukocyte poor I think 80% is good enough platelet recovery. The WBC in the protocol shown usually drops from baseline as opposed to a LR-PRP where I would get 1 or 2 cc of the "red" which would give more platelets and leukocytes also.