Method for manufacture of macrobeads

Abstract

The invention relates to an improved method for making agarose coated, agarose beads which contain cells. The method which is preferably automated, involves placing manufactured beads in a sample of mineral oil at a temperature gradient, such that the temperature drops as the bead moves through the oil. Preferably, a “trumpet tool” and a “straw tool” are employed in the method.

We claims: 
     
       1. A method for producing a composition of matter comprising a sample of live cells in an agarose containing bead, wherein said bead is coated with agarose, comprising:
 (a) mixing a first sample of agarose with said sample of live cells to form a suspension; 
 (b) moving said suspension to a first sample of mineral oil, to form a bead from said suspension; 
 (c) removing said bead from said first sample of mineral oil; 
 (d) rinsing mineral oil from said bead; 
 (e) moving said bead with a trumpet tool to a second solution of agarose; 
 (f) coating said bead with said second solution of agarose, 
 (g) removing said bead from said second solution of agarose with a straw tool, and 
 (h) dispensing said coated bead into a second sample of mineral oil, wherein said sample of mineral oil is kept at a temperature gradient so that said bead moves along a path from a location in the mineral oil that is at a higher temperature of about 20° C. to about 30° C. to a location in the mineral oil at a lower temperature of about 0° C. to about −8° C. 
 
     
     
       2. The method of  claim 1 , wherein said cells are secretory cells. 
     
     
       3. The method of  claim 1 , wherein said cells are cancer cells. 
     
     
       4. The method of  claim 1 , wherein said cells are cancer stem cells. 
     
     
       5. The method of  claim 1 , wherein said cells are islet cells. 
     
     
       6. The method of  claim 1 , wherein said cells are stem cells. 
     
     
       7. The method of  claim 6 , wherein said cells are embryonic stem cells. 
     
     
       8. The method of  claim 1 , wherein said cells are pluripotent cells. 
     
     
       9. The method of  claim 1 , further comprising removing said bead from said first or second sample of mineral oil with a trumpet tool. 
     
     
       10. The method of  claim 1 , wherein said higher temperature is from about 20° C. to about 25° C. and said lower temperature is from about 0° C. to about −2° C. 
     
     
       11. The method of claim 1, wherein said first sample of agarose has a gel strength of ≥1000 g/cm 2  at a 1.5% gel, and a viscosity of from 5.8 to 8.7 cP, a gelling temperature of 40-43° C. for a 1.5% solution, an electro endosmosis value of from 0.0 to 0.12, and a sulphate content ≤0.30%. 
     
     
       12. The method of claim 1, wherein said second sample of agarose has a gel strength of ≥1000 g/cm 2  at a 1.5% gel, and a viscosity of from 5.8 to 8.7 cP, a gelling temperature of 40-43° C. for a 1.5% solution, an electroendosmosis value of from 0.0 to 0.12, and a sulphate content ≤0.30%. 
     
     
       13. The method of claim 1, wherein said first and second samples of agarose have a gel strength of ≥1000 g/cm 2  at a 1.5% gel, and a viscosity of from 5.8 to 8.7 cP, a gelling temperature of 40-43° C. for a 1.5% solution, an electroendosmosis value of from 0.0 to 0.12, and a sulphate content ≤0.30%. 
     
     
       14. The method of claim 1, wherein said live cells are cancer cells. 
     
     
       15. The method of claim 14, wherein said cancer cells are renal cancer cells. 
     
     
       16. The method of claim 11, wherein said live cells are islets. 
     
     
       17. The method of claim 11, wherein said live cells are stem cells.