Chemistry: molecular biology and microbiology – Animal cell – per se ; composition thereof; process of...
Reexamination Certificate
2000-09-20
2001-08-07
Redding, David A. (Department: 1744)
Chemistry: molecular biology and microbiology
Animal cell, per se ; composition thereof; process of...
C435S383000, C435S395000, C435S286500, C435S286600, C435S294100, C435S305200
Reexamination Certificate
active
06271027
ABSTRACT:
The invention relates to the field of cell and tissue culture with the help of a culture fluid or nutrient medium.
This invention more particularly relates to devices and methods for cell and tissue culture in which the culture fluid (or nutrient) is set into motion so as to achieve a dynamic culture. Such devices usually apply either a technique for directly stirring the culture fluid in the culture volume or a technique providing permanent flow of the fluid through a specific circuit.
The present invention is more specifically directed to fluid flow devices which comprise at least a culture well defining a chamber in which cells or tissues to be grown are accommodated, first and second tanks, at least one of which is configured so as to receive a culture fluid, as well as connecting means coupling the well to the first and second tanks. The culture fluid flows from one tank to the other via the well.
This technique for fluid flow is difficult to control because culture conditions change over time. Actually, the low cell concentration which exists at the beginning of the culture process, requires a quasi-static environment, and therefore a very low culture fluid flow rate. Then the flow rate rapidly increases because of cell multiplication or tissue growth.
Maintaining adequate conditions during the whole culture process requires either the use of several suitable devices, at different stages of the culture's growth, respectively, and this involves many handling operations, or complex and often cumbersome means, which limit, even preclude observation of the culture's development with a microscope. These handling operations and means are all the more complex as the culture process should be carried out in a sterile environment which requires highly qualified personnel, and this therefore increases the cost for each culture. Furthermore, because of their price, these devices should be reusable, which imposes complete sterilization before each new culture, and maintenance operations.
The object of the invention is therefore to overcome all or part of the aforementioned drawbacks.
For this purpose, it provides a device of the type described in the introduction wherein each of the first and second tanks accommodates at least a flexible pocket, wherein at least one of the pockets of these tanks is adapted for receiving the culture fluid, and which comprises at least a control means able to provide first and/or second sequences of external pressures to be applied, on the pockets of the first and second tanks, respectively, and pressurization means configured for applying to the pockets, whereby the pressures are defined by the first and second sequences provided by the controller module.
So, defining for each type of culture, pressure sequences to be applied on the different pockets, then transferring these sequences to the controller module is sufficient for carrying out the culture process throughout its duration in adequate conditions. This transfer may indirectly be made through an entry interface, or else directly through a memory in which a multiplicity of sequences matching different samples, is stored beforehand (this memory may possibly be rewritable through the aforementioned interface).
In a preferred embodiment of the device according to the invention, the first and second tanks each comprise an upper portion and a lower portion which are connected to each other through an intermediate narrow portion and each accommodate a flexible pocket, wherein the upper and lower flexible pockets communicate with each other via the intermediate portion, and connecting means communicating with the lower pockets.
In this embodiment, it is particularly advantageous that the controller module be able to provide pressure sequences suitable for each pocket, so that the pressurization means apply on each of these pockets the pressures from the sequences that match them.
The device according to the invention may include additional characteristics considered separately or combined and notably:
At least one of the first and second tanks may include an opening adapted for introducing culture fluid into the communication duct between pockets;
at least one of the upper portions of the tanks may include an upper opening for connecting the upper pocket, which it accommodates, to a culture fluid feeding and extraction device;
first access control means, driven by the controller module may control access to the upper pocket via the upper port;
each lower tank portion may include a lower port for connection of connecting means to the lower pocket which it accommodates, and second access control means, driven by the controller module, may control access to the lower pockets via the lower ports;
the upper and lower portions of the first and second tanks may each include a leak-proof inlet, and the pressurization means may comprise a first pressurization circuit supplying with high pressure fluid, under control of lower and upper valves driven by the controller module, each upper and lower portion of the first and second tanks, via the leak-proof inlets;
the upper and lower portions of the first and second tanks may each include a leak-proof inlet, and the pressurization means may comprise, on the one hand, a first pressurization circuit including a fluid pump (or compressor) and supplying with high pressure fluid, under the control of upper and lower valves driven by the controller module, each upper and lower portion of the first tank via the leak-proof inlets, and on the other hand, a second pressurization circuit including a fluid pump and supplying with high pressure fluid, under the control of upper and lower valves driven by the controller module, each upper and lower portion of the second tank, via the leak-proof inlets;
each pressurization circuit may also comprise two other valves respectively cooperating with the first and second access control means, which are then advantageously provided with a duct including a flexible thinned area on which the high pressure fluid may act
the pressurization means may also include a third pressurization circuit supplying with low pressure fluid, under the control of upper and lower valves, each upper and lower portion of the first and second tanks, via leak-proof inlets. Alternatively the pressurization means may include a third pressurization circuit supplying with low pressure fluid, under the control of said upper and lower valves, each upper and lower portion of the first tank, via the leak-proof inlets, as well as a fourth pressurization circuit supplying with low pressure fluid, under the control of upper and lower valves, each upper and lower portion of the second tank, via the leak-proof inlets. In each case, each third and fourth pressurization circuit is advantageously connected to the other valves so as to cooperate with the first and second access control means;
at least two wells may be placed in series by having them communicate through connecting means, wherein each of these wells is connected to the first tank and the other well is connected to the second tank. A third or even a fourth or fifth well may be placed in series between the two other wells;
the wells may be provided as independent units assembled together, or they may be configured on a culture plate;
each culture chamber may include at least a transparent wall for observation of cells and tissues during the culture process, for example under a microscope;
each well may be configured so as to define two superimposed culture chambers separated by a filter.
In addition, the invention relates to a cell and tissue culture facility comprising at least two devices of the type described above, placed in parallel, and including a single control unit driving together the control units of these devices.
The invention also relates to a cell and tissue culture method. This method starts with a known first step wherein a culture device is provided including at least a culture well for receiving the cells or tissues to be grown, with first and second tanks coupled to the well, at leas
Sarem Farzin
Sarem Damerdji Leila-Ouassila
Cell Tissue Progress
Redding David A.
Young & Thompson
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