Platelet rich plasma formulations and use thereof

Abstract

Compositions for platelet rich plasma (PRP), depleted in neutrophils, are provided. Generally, these compositions comprise a higher concentration of platelets, lymphocytes and monocytes than whole blood with selective depletion of neutrophils to a concentration of less than about 5000/μl. These compositions may have depressed concentrations of red blood cells and hemoglobin. In some variations, the compositions may be useful to treat damaged connective tissue and/or to slow or stop cardiac apoptosis after a heart attack. The PRP composition may be delivered in conjunction with reperfusion therapy.

What is claimed is: 
     
         1 . A method of treating damaged connective tissue comprising:
 delivering a composition to the patient to treat the damaged connective tissue comprising:   platelets in a concentration of about 500,000/μl to about 7,000,000 μl,   monocytes in a concentration of about 400/μl to about 3200/μl,   neutrophils in a concentration of less than about 5000/μl, and   lymphocytes in a concentration of about 2600/μl to about 16000/μl, wherein the composition does not include an exogenous activator.   
     
     
         2 . The method of  claim 1 , further comprising preparing the composition from the whole blood of the patient. 
     
     
         3 . A method of preparing platelet-rich plasma (PRP) from a patient comprising:
 withdrawing blood from the patient;   preparing PRP from the blood, and   administering reperfusion therapy to the patient,   
       wherein a thrombolytic agent has not been administered to the patient prior to withdrawing the blood, and wherein the PRP comprises:
 platelets in a concentration of about 500,000/μl to about 7,000,000 μl, 
 monocytes in a concentration of about 400/μl to about 3200/μl, 
 neutrophils in a concentration of less than about 5000/μl, and 
 lymphocytes in a concentration of about 2600/μl to about 16000/μl. 
 
     
     
         4 . The method of  claim 3 , wherein the thrombolytic agent is selected from the group consisting of heparin, tPA, PLAVIX®, and aspirin. 
     
     
         5 . The method of  claim 3 , wherein the patient has not received a thrombolytic agent for at least 2 hours, preferably 1 day, more preferably 2 weeks, and yet more preferably 1 month prior to withdrawing the blood. 
     
     
         6 . The method of  claim 3 , wherein preparing PRP from the blood comprises the steps of:
 obtaining a plasma fraction from the blood;   isolating platelets from the plasma fraction;   resuspending the platelets in a reduced amount of plasma; and   adjusting the pH to provide a pH of 7.3 to 7.5 for the resuspended platelets to provide a platelet-rich plasma composition,   
       wherein an activator of the platelet-rich plasma is not added to the platelet-rich plasma composition. 
     
     
         7 . A method of identifying drug candidates for treating a disease or condition comprising administering a platelet rich plasma composition to an individual suffering from a disease or condition or a disease model of that disease selected from the group consisting of a cell culture, computer model and animal model;
 monitoring the efficacy of the treatment in the individual suffering from the disease or condition or in the model of that disease;   selecting for individual or results from the model responsive to the treatment;   obtaining a blood, saliva or tissue sample from the responsive individuals or responsive models;   analyzing the samples for a predetermined marker(s);   comparing the analysis of the samples to individuals or models of the disease not responsive to the treatment; and   identifying analytes which are responsive to the treatment; and   screening for drugs affecting the analytes, thereby identifying drugs candidates for treating the disease or condition.   
     
     
         8 . The method of  claim 7 , wherein the individuals or models not responsive to the treatment do not have the disease or condition. 
     
     
         9 . The method of  claim 7 , wherein the individuals or models not responsive to the treatment have the disease or condition but are not responsive to the treatment. 
     
     
         10 . The method of  claim 7 , wherein the disease or condition is selected from the group consisting of ischemia, cancer, a disease of the immune system, a connective tissue injury, a skin disease, and a disease of the nervous system. 
     
     
         11 . The method of  claim 10  wherein the disease or condition is ischemia and wherein the ischemia is cardiac ischemia. 
     
     
         12 . The method of  claim 10 , wherein the disease or condition is cancer and the cancer is brain cancer. 
     
     
         13 . The method of  claim 10 , wherein the disease or condition is a connective tissue injury which is a tendinosis of elbow, shoulder, or knee. 
     
     
         14 . The method of  claim 10 , wherein the disease or condition is a disease of the nervous system which is Parkinsons' disease. 
     
     
         15 . The method of  claim 7 , wherein the treatment comprises administering a PRP composition to a tumor, cardiac tissue, or connective tissue. 
     
     
         16 . The method of  claim 7 , wherein the PRP composition comprises neutrophils at a level of 0 to 0.9 of the concentration of whole blood. 
     
     
         17 . The method of  claim 7 , wherein the analysis comprises determining a genetic profile. 
     
     
         18 . The method of  claim 7 , wherein the analysis comprises measuring levels of proteins, lipids, antibodies, antigens, enzymes or small molecules. 
     
     
         19 . A method of disease diagnosis for a patient comprising:
 obtaining a blood sample from the patient;   obtaining PRP from the blood sample;   obtaining an analysis from the PRP; and   comparing the analysis of the PRP from the patient with an analysis obtained from PRP of a population having the disease; and   determining that analysis of the PRP of the patient matches the analysis of the PRP of the disease population, thereby diagnosing the disease,   wherein the analysis is an immunoassay or genetic profile.   
     
     
         20 . A device comprising a platelet rich plasma composition comprising:
 platelets in a concentration of about 500,000/μl to about 7,000,000/μl,   monocytes in a concentration of about 400/μl to about 3200/μl,   neutrophils in a concentration of less than about 5000/μl, and   lymphocytes in a concentration of about 2600/μl to about 16000/μl, wherein the composition does not include an exogenous activator optionally in combination with a fixation device selected from the group consisting of a stent, suture, screw and implantable device.   
     
     
         21 . The device according to  claim 20 , wherein the device is a chamber and wherein conditions in the chamber include one or more of the following: low oxygen tension, high oxygen tension, low pH, high pH, low pressure, high pressure, low UV, high UV, low temperature and high temperature.