Method and apparatus for maintaining microcarrier beads in suspension
View Patent ↗The invention relates to an impeller for use in a bioreactor for growing adherent mammalian cells and/or a cell separator for adherent mammalian cells. The impeller of the invention is designed to efficiently and rapidly lift a bed of settled cells and microcarrier beads at low rotational speeds. Cell growth and concomitant yield is therefore improved by maintaining the culture in an environment of low shear force.
1. An impeller for use in a bioreactor for adherent mammalian cells and/or a cell separator for adherent mammalian cells comprising:
a hub attachable to a rotatable shaft;
one or more blades attached to said hub;
each said blade comprising:
a first side adjacent to the hub;
a second side and a third side adjacent to and at right angles to said first side, said second and third sides defining a height therebetween;
a fourth side opposed to the first side and forming an outer edge of the blade, said outer edge disposed at a length from said hub; and
a surface having a planar portion and a curved portion;
wherein said curved portion is angled in the range of 70 to 120° to the plane of said planar portion to form a leading surface relative to the direction of rotation of said impeller,
wherein the second side is shorter than the third side,
wherein the impeller is configured to turn the mammalian cells at low shear force to minimize cellular injury,
wherein the fourth side outer edge includes a generally straight portion merging into said curved portion; and said length being greater than said height.
2. The impeller of claim 1 , wherein the curved portion is angled in the range of 80 to 100° to the plane of the curved portion.
3. The impeller of claim 1 , wherein the curved portion is angled at 90° to the plane of the planar portion ( 36 ).
4. The impeller of claim 1 , wherein the curved portion constitutes 20 to 40% of the area of said surface.
5. The impeller of claim 1 , wherein the curved portion is curved upwards.
6. A method for growing adherent mammalian cells, said method comprising:
a) adding adherent mammalian cells to growth medium in a bioreactor;
b) stirring said growth medium with the impeller of claim 1 ;
wherein the impeller is rotated at a speed of 10 to 55 rpm.
7. The method of claim 6 , wherein said cells are attached to microcarrier beads prior to adding the cells to said growth medium in said bioreactor.
8. The method of claim 6 , wherein the growth medium contains microcarrier beads and the cells attach to said beads while the impeller is stirring the growth medium.
9. The method of claim 6 , wherein the impeller is rotated at a speed of 15 to 30 rpm.
10. A bioreactor or a cell separator comprising the impeller of claim 1 .
11. The bioreactor or cell separator of claim 10 , further comprising a head plate, a cylindrical wall, and a base, wherein said cylindrical wall is secured to said head plate and said base.
12. The cell separator of claim 11 , further comprising:
a cylindrical sieve attached to the head plate and enclosing the impeller to form an inner chamber for separating cells from microcarrier beads within and an outer chamber for collecting cells therefrom;
an inlet for feeding cells and microcarrier beads into the inner chamber; and
an outlet for collecting cells from the outer chamber;
wherein the sieve comprises a first mesh having a plurality of pores of a size that permits the flow of cells through to said outer chamber but excludes the passage of microcarrier beads.
13. The cell separator of claim 12 , wherein said sieve additionally comprises a second mesh having a plurality of pores of a size greater than that of said pores of said first mesh.
14. The cell separator of claim 12 , further comprising a sparger attached to said inlet.