Method for Obtaining Sterile Human Amniotic Fluid and Uses Thereof

Abstract

Provided herein is a pre-Caesarean method for collecting amniotic fluid from a patient. A needle is inserted into the incision site for the future C-section, which may be under ultrasound guidance, through which the amniotic fluid is drawn under a low level suction and, optionally, gravity to a sterile collection container. The method encompasses filtering and/or irradiating the amniotic fluid to collect biomolecules of interest such as growth factors and/or stem cells. Also provided is the sterile amniotic fluid or filtrates thereof collected by the method described herein

What is claimed is: 
     
         1 . A method of obtaining sterile human amniotic fluid from an individual, comprising the steps of:
 inserting a blunt tip needle into the amniotic sac of said individual;   attaching said blunt tip needle to a three-way stopcock;   connecting a Luer lock syringe to said three-way stopcock;   connecting a first end of a length of sterile tubing with said three-way stopcock; and   collecting sterilely said amniotic fluid through the blunt tip needle and sterile tubing into a collection container.   
     
     
         2 . The method of  claim 1 , wherein collecting of said amniotic fluid is performed under ultrasound guidance. 
     
     
         3 . The method of  claim 1 , wherein said sterile collection container comprises a pump with a suction device. 
     
     
         4 . The method of  claim 3 , wherein said suction device is a low suction device or spring loaded low suction device. 
     
     
         5 . The method of  claim 3 , wherein said suction device is fluidly connected to an internal balloon. 
     
     
         6 . The method of  claim 5 , further comprising manually pumping up the internal balloon in the sterile collection container using the low suction device to allow a low-level suction and collection of the amniotic fluid. 
     
     
         7 . The method of  claim 3 , wherein said sterile collection container comprises an inlet. 
     
     
         8 . The method of  claim 7 , further comprising connecting a second end of the tubing to said inlet of the sterile collection container. 
     
     
         9 . The method of  claim 3 , wherein said sterile collection container comprises a vent having a cap. 
     
     
         10 . The method of  claim 9 , wherein said vent is resealed after full expansion of the internal balloon. 
     
     
         11 . The method of  claim 10 , further comprising sterilizing said amniotic fluid by filtration. 
     
     
         12 . The method of  claim 11 , wherein said sterilizing step comprises filtering said amniotic fluid through 100 μm filters to remove large particles while retaining stem cells to obtain prefiltered amniotic fluid. 
     
     
         13 . The method of  claim 11 , wherein said sterilizing step comprises passing prefiltered amniotic fluid through a filter having a size of from about 10 μm to about 1 μm so to remove cells and large particles to obtain a micron filtrate. 
     
     
         14 . The method of  claim 13 , further comprising passing the micron filtrate through a filter having a size of about 0.45 μm to obtain a submicron filtrate. 
     
     
         15 . The method of  claim 14 , further comprising irradiating the submicron filtrate to obtain an irradiated filtrate. 
     
     
         16 . The method of  claim 14 , wherein said irradiation is by e-beam irradiation or gamma ray irradiation. 
     
     
         17 . The method of  claim 15 , further comprising passing the irradiated filtrate through a filter having a size of about 0.22 μm to achieve sterile amniotic fluid. 
     
     
         18 . Sterile amniotic fluid prepared by the method of  claim 1 . 
     
     
         19 . The sterile amniotic fluid of  claim 18 , said fluid comprising a filtrate containing biomolecules selected via filtration. 
     
     
         20 . The sterile amniotic fluid of  claim 19 , wherein said biomolecules are growth factors or stem cells or a combination thereof.