Microencapsulation process of sertoli cells, microcapsules obtained and use for treatment of type i diabetes mellitus

Abstract

An apparatus for production of microcapsules includes a volumetric pump configured to deliver a polysaccaridic suspension through a catching tube. A needle-type element is configured to receive the suspension through the catching tube, and the needle-type element has a button hole opening in a lateral wall thereof and an output hole that outputs the suspension. The button hole opening receives a pressure fluid current. A pressure regulator is coupled to the button hole opening to regulate the pressure fluid current to interrupt a suspension flow and obtain microdroplets of homogeneous size exiting the output hole. The microdroplets are received in a receiving container in a solution including divalent cations or polycationic substances to form a gel such that homogeneous microcapsules are formed.

1 .- 13 . (canceled) 
     
     
         14 . An apparatus for production of microcapsules, comprising:
 a volumetric pump configured to deliver a polysaccaridic suspension through a catching tube;   a needle-type element configured to receive the suspension through the catching tube, the needle-type element having a button hole opening in a lateral wall thereof and an output hole that outputs the suspension, the button hole opening receiving a pressure fluid current;   a pressure regulator coupled to the button hole opening to regulate the pressure fluid current to interrupt a suspension flow and obtain microdroplets of homogeneous size exiting the output hole, the microdroplets being received in a receiving container in a solution including divalent cations or polycationic substances to form a gel such that homogeneous microcapsules are formed.   
     
     
         15 . The apparatus as recited in  claim 14 , wherein the polysaccaridic suspension includes a homogeneous suspension of Sertoli Cells (SC) with purity, in terms of cell composition, higher than 90%, in a saline solution of ultrapure sodium alginate. 
     
     
         16 . The apparatus as recited in  claim 15 , wherein the needle-type element extrudes the suspension through the output hole to obtain a continuous flow of microdroplets showing homogeneous dimensions. 
     
     
         17 . The apparatus as recited in  claim 16 , wherein the homogeneous dimensions include the microcapsules with substantially fixed diameters and without tails structures. 
     
     
         18 . The apparatus as recited in  claim 16 , wherein the homogeneous dimensions include the microdroplets having a mean diameter of between about 300 and 700 microns with a standard deviation below 40 microns. 
     
     
         19 . The apparatus as recited in  claim 14 , wherein the microcapsules include sodium alginate, at a concentration of 1-3% w/v, with an endotoxin content not exceeding 20 EU/g and a protein content lower than 0.4%. 
     
     
         20 . The apparatus as recited in  claim 14 , further comprising a fluid generator coupled to a supply side of the pressure regulator and having a pressure reduction to obtain a pressure fall of at least 0.3 bars in a flow/no flow transient of fluid current. 
     
     
         21 . The apparatus as recited in  claim 19 , wherein the fluid generator includes a linear relationship between pressure cycles and amount of fluid dispensed. 
     
     
         22 . The apparatus as recited in  claim 14 , wherein the microcapsules each include at least 20×10 6  Sertoli Cells (SC). 
     
     
         23 . The apparatus as recited in  claim 14 , wherein the microcapsules including the Sertoli Cells are suitable as a sole therapeutic agent for the prevention and treatment of Type 1 diabetes mellitus (T1DM). 
     
     
         24 . An apparatus for production of microcapsules including Sertoli Cells, comprising:
 a volumetric pump configured to deliver a polysaccaridic suspension through a catching tube, wherein the polysaccaridic suspension includes a homogeneous suspension of Sertoli Cells with purity, in terms of cell composition, higher than 90%, in a saline solution of ultrapure sodium alginate;   a needle-type element configured to receive the suspension through the catching tube, the needle-type element having a button hole opening in a lateral wall thereof and an output hole that outputs the suspension, the button hole opening receiving a pressure fluid current wherein the needle-type element extrudes the suspension through the output hole to obtain a continuous flow of microdroplets showing homogeneous dimensions;   a pressure regulator coupled to the button hole opening to regulate the pressure fluid current to interrupt a suspension flow and obtain microdroplets of homogeneous size exiting the output hole, the microdroplets being received in a receiving container in a solution including divalent cations or polycationic substances to form a gel such that homogeneous microcapsules are formed; and   a fluid generator coupled to a supply side of the pressure regulator and configured to supply the pressure fluid current.   
     
     
         25 . The apparatus as recited in  claim 23 , wherein the homogeneous dimensions include the microcapsules with substantially fixed diameters and without tails structures. 
     
     
         26 . The apparatus as recited in  claim 24 , wherein the homogeneous dimensions include the microdroplets having a mean diameter of between about 300 and 700 microns with a standard deviation below 40 microns. 
     
     
         27 . The apparatus as recited in  claim 24 , wherein the microcapsules include sodium alginate, at a concentration of 1-3% w/v, with an endotoxin content not exceeding 20 EU/g and a protein content lower than 0.4%. 
     
     
         28 . The apparatus as recited in  claim 24 , wherein the fluid generator has a pressure reduction applied to obtain a pressure fall of at least 0.3 bars in a flow/no flow transient. 
     
     
         29 . The apparatus as recited in  claim 28 , wherein the fluid generator includes a linear relationship between pressure cycles and amount of fluid dispensed. 
     
     
         30 . The apparatus as recited in  claim 24 , wherein the microcapsules each include at least 20×10 6  Sertoli Cells (SC). 
     
     
         31 . The apparatus as recited in  claim 24 , wherein the microcapsules including the Sertoli Cells are suitable as a sole therapeutic agent for the prevention and treatment of Type 1 diabetes mellitus (T1DM). 
     
     
         32 . An apparatus for production of microcapsules including Sertoli Cells, comprising:
 a container including a homogeneous suspension of Sertoli Cells with a higher than 90% purity, in terms of cell composition, in a saline solution of ultrapure sodium alginate;   a volumetric pump configured to aspirate the suspension to create a suspension flow;   a needle-type element configured to receive the suspension flow;   a pressure regulator coupled to the needle-type element and configured to regulate a pressure fluid current therethrough to interrupt the suspension flow and obtain microdroplets of homogeneous size exiting an output hole of the needle-type element, the microdroplets being received in a receiving container in a solution including divalent cations or polycationic substances to form a gel such that homogeneous microcapsules are formed; and   a fluid generator coupled to a supply side of the pressure regulator and configured to supply the pressure fluid current.   
     
     
         33 . The apparatus as recited in  claim 32 , wherein the microcapsules have substantially fixed diameters without tails structures and have a mean diameter of between about 300 and 700 microns with a standard deviation below 40 microns.