Process for the identification and isolation of insulin producing beta cell differentiating factor from the perivitalline fluid of the fertilised eggs of horseshoe crab

Abstract

This invention relates to the identification and characterization of cell proliferating factor in the peri-vatalline fluid of the fertilized eggs of the Indian horseshoe crab.

We claim  
     
         1 . A process for identification of insulin producing β-cells proliferating factor from the peri-vitalline fluid of fertilized eggs of horseshoe crab that facilitates the proliferation of AR42J cells from rat origin.  
     
     
         2 . A process as claimed in  claim 1  wherein the process comprises collecting peri-vitalline fluid from the fertilized eggs of the Indian horseshoe crab  
     
     
         3 . A process as claimed in  claim 1  wherein the peri-vitalline fluid is collected aseptically, aliquated and stored at −20° C.  
     
     
         4 . A process as claimed in  claim 1  wherein the AR42J cell line (rat ascinar cells) obtained from American Type Culture Collection (Rockville Md, U.S.A.) was grown in Dulbeco's modified minimum essential medium supplemented with 10% FCS.  
     
     
         5 . A process as claimed in  claim 1  wherein the cells were maintained in NUNC six-well dishes (NUNCLON, Denmark).  
     
     
         6 . A process as claimed in  claim 1  wherein the a sub culture is done every 4 th  day of seeding of 1×10 5  cells and the cultures were incubated at 37° C.  
     
     
         7 . A process as claimed in  claim 1  wherein the concentration dependent proliferation was observed in presence of peri-vitalline fluid.