Centrifuge Fermenter Array

Abstract

A centrifuge microplate fermenter for culturing cells in a nutrient medium wherein the centrifuge microplate fermenter is comprised of a top end, a bottom end, a vertical cylindrical outer wall positioned between the top end and the bottom end, and an inner wall positioned inside the outer wall, the bottom end further comprised of a plurality of baffles which protrude from inner wall toward a theoretical center of the vertical cylindrical outer wall terminating in a conical tip

Claims

1. A centrifuge microplate fermenter with plurality of wells for culturing cells in a nutrient medium, the centrifuge microplate fermenter comprised of: a top end a bottom four side walls a plurality of wells each well having a vertical cylindrical outer wall positioned between the top end and the bottom end; and an inner wall positioned inside the outer wall, wherein the bottom end is further comprised of a plurality of baffles which protrude from inner wall toward a theoretical center of the vertical cylindrical outer wall terminating in a conical tip.

2. The centrifuge microplate fermenter of claim 1, wherein the centrifuge microplate fermenter is fabricated from an appropriate material selected from the group consisting of polypropylene/polysulfone, polyethersulfone, ABS/polycarbonate, poluolefin, polyethylene and polyacrylic plastic.

3. The centrifuge microplate fermenter of claim 1, wherein the centrifuge microplate fermenter is fabricated by injection molding.

4. A centrifuge microplate fermenter for culturing cells in a nutrient medium, the centrifuge microplate fermenter comprised of: a top end, a bottom end; four side walls and a plurality of wells each having a vertical cylindrical outer wall positioned between the top end and the bottom end; and an inner wall positioned inside the outer wall, wherein the bottom end is further comprised of a plurality of baffles which protrude from inner wall toward a theoretical center of the vertical cylindrical outer wall terminating in a conical tip.

5. The centrifuge microplate fermenter of claim 1, wherein there are a plurality of (2-4) individual baffles that extend upward from the conical tip - the length of the vertical cylindrical inner wall (see FIG. 4, 8-1).

6. The centrifuge microplate fermenter of claim 5, wherein each individual baffle starts in the approximate middle of the conical tip and tapers vertically and horizontally up the inner wall toward the top (open) end to a point of termination (see FIG. 4, 8-1).

7. The centrifuge microplate fermenter of claim 5, wherein the plurality of baffles take up approximately - of the vertical cylindrical inner wall radial diameter (see FIG. 5, 8-1, 8-2, 8-3, 8-4)

8. The centrifuge microplate fermenter of claim 1, wherein the centrifuge microplate fermenter is fabricated with a total volume selected from the range of about 250 ul to about 50 ml.

9. A method for improving cell culturing, the method comprising: providing a centrifuge microplate fermenter with side and bottom baffles terminating in a conical tip; placing cell culture media and one or more inocula cells inside the centrifuge microplate fermenter; allowing one or more cells to divide in the media to produce progeny cells; centrifuging the centrifuge fermenter tube; and removing the cell culture media by aspiration thereby leaving the cultured cell pellet for further use.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0016] The novel features of this invention, as well as the invention itself, both as to its structure and its operation, will be best understood from the accompanying drawings, taken in conjunction with the accompanying description, in which similar reference characters refer to similar parts, and in which:

[0017] FIG. 1 illustrates a longitudinal cross section view of the centrifuge microplate fermenter.

[0018] FIG. 2 illustrates a top view of the centrifuge microplate fermenter of FIG. 1.

[0019] FIG. 3 illustrates a longitudinal cross section view of a single well of the centrifuge microplate fermenter of FIG. 1.

[0020] FIG. 4 illustrates a magnified cross section view of the encircled bottom of the centrifuge microplate fermenter of FIG. 1.

[0021] FIG. 5 illustrates a magnified top view of the of the centrifuge microplate fermenter of FIG. 1.

[0022] FIG. 6 illustrates the results of a cell growth comparison study performed using the centrifuge microplate fermenter and standard 15 ml cell culture centrifuge microplate.

DETAILED DESCRIPTION OF THE INVENTION

[0023] A centrifuge microplate fermenter 1 for culturing cells in a nutrient medium is shown in FIG. 1. Centrifuge microplate fermenter is comprised of a plurality of wells 2 as shown in FIG. 2. Centrifuge microplate fermenter consists of a top end 3 and a bottom end 4, the top end 3 and the bottom end 4 terminating in a conical tip 9

Centrifuge microplate fermenter is further comprised of a vertical cylindrical wall 5 positioned between top end 3 and bottom end 4. Vertical cylindrical wall 5 is further comprised of an inner wall 6 and outer wall 7. Bottom end 4 is further comprised of a plurality of baffles 8-1 which protrude from inner wall 6 toward the theoretical center of the vertical cylindrical wall 5.

[0024] With reference to FIG. 4, a magnified view of the bottom end of the centrifuge microplate fermenter 1 shown in FIG. 1 is illustrated. The conical tip 9 is visible beneath the plurality of baffles 8-1 as is the theoretical center 10 of the vertical cylindrical wall 5.

[0025] With reference to FIG. 5, a top view of the centrifuge microplate fermenter of FIG. 1 is illustrated. The position of the plurality of baffles (8-1, 8-2, 8-3, 8-4) is shown in relation to the theoretical center 10 of the vertical cylindrical wall (not shown). A plurality of baffles equals four baffles for the 2 ml centrifuge microplate fermenter as shown. The number of baffles may be increased or decreased according to the volume of the centrifuge microplate and the amount of increased aeration desired where the number of baffles will be at least one.

Example 1

[0026] Small-scale preparation of plasmid DNA was performed in the centrifuge microplate fermenter of the present invention and a standard culture centrifuge microplate to compare cell growth/plasmid yield.

Methods

[0027] A single bacterial colony was transferred into 0.5 ml of amp LB medium, containing 50 g/ml ampicillin in multiple wells of 2.0 ml centrifuge microplate fermenter and standard microplate. Cell cultures were incubated overnight at 37 C. with vigorous shaking. Culture centrifuge microplates were centrifuged at 12,000 g for 30 seconds at 4 C. The supernatant was removed by aspiration and the cells in the bacterial pellet were counted and compared.

[0028] FIG. 6 shows the results of the DNA yield comparison study. DNA yield of cultures grown in standard microplate are shown in the front row while the DNA yield of cultures grown in the centrifuge microplate fermenter of the present invention are shown in the back row. The DNA yield in the centrifuge microplate fermenter increased by over 60% compared to the standard microplates. These results confirm that the design of the centrifuge microplate fermenter drastically increases bacterial culture growth resulting in a 60% increase in DNA yield per unit volume.

[0029] One embodiment of the present invention provides a centrifuge microplate fermenter for culturing cells in a nutrient medium, the centrifuge microplate fermenter comprised of:

[0030] a top end

[0031] a bottom end;

[0032] a vertical cylindrical outer wall positioned between the top end and the bottom end; and

[0033] an inner wall positioned inside the outer wall,

[0034] wherein the bottom end is further comprised of a plurality of baffles which protrude from inner wall toward a theoretical center of the vertical cylindrical outer wall

[0035] In another embodiment of the present invention, the centrifuge microplate fermenter can be fabricated from any appropriate material, including but not limited to, polypropylene/polysulfone, polyethersulfone, AB S/polycarbonate, a polyacrylic plastic and/or the like. In a preferred embodiment, the centrifuge microplate fermenter is injection molded from virgin material polyolefin, such as high-density polypropylene or polyethylene. In a most preferred embodiment, the centrifuge microplate fermenter is injection molded from ultra-clear medical grade polypropylene.

[0036] In yet another embodiment of the present invention, the centrifuge microplate fermenter may be fabricated in a variety of volumes ranging from 0.6 ml to 50 ml. In a preferred embodiment, the centrifuge microplates are centrifuge microplates with a volume ranging from 250 l to 2 ml. In a preferred embodiment, the centrifuge microplates can be arranged and molded into 6, 12, 24, 48, 96, 192, 288, 384 well array configurations. In the most preferred red embodiment, the micro-centrifuge centrifuge microplate has a volume of 1-2 ml and can be arranged and molded in a 96 well array.

[0037] It will be appreciated that details of the foregoing embodiments, given for purposes of illustration, are not to be construed as limiting the scope of the invention. Although several embodiments of this invention have been described in detail above, those skilled in the art will readily appreciate that many modifications are possible in the exemplary embodiments without materially departing from the novel teachings and advantages of this invention. Accordingly, all such modifications are intended to be included within the scope of this invention, which is further defined in the converted utility application and appended claims. Further, it is recognized that many embodiments may be conceived that do not achieve all the advantages of some embodiments, particularly preferred embodiments, yet the absence of advantage shall not be construed to necessarily mean that such an embodiment is outside the scope of the present invention.