Bioreactors with substance injection capacity

A bioreactor comprising:

• a. a first substrate having a first surface, an opposite second surface and edges;
• b. a second substrate having a first surface and an opposite second surface, defining a cavity with a bottom surface, wherein the bottom surface is located therebetween the first surface and the second surface, and wherein the first surface of the first substrate is received by the second surface of the second substrate to cover the cavity so as to form a chamber for receiving cells and a liquid medium;
• and
• c. perfusion means in fluid communication with the chamber to allow diffusional exchange of nutrients and metabolic byproducts with the chamber.

A bioreactor comprising:

• a. a first substrate having a first surface, an opposite second surface and edges;
• b. a second substrate having a first surface and an opposite second surface, defining a cavity with a bottom surface, wherein the bottom surface is located therebetween the first surface and the second surface, and wherein the first surface of the first substrate is received by the second surface of the second substrate to cover the cavity so as to form a chamber for receiving cells and a liquid medium;
• c. a filter dividing the chamber into a first subchamber and a second subchamber, wherein the filter has a porosity to allow the first subchamber and the second subchamber in fluid communication;
• and
• d. a port formed between the bottom surface and the first surface of the second substrate with a first opening and an opposite, second opening, wherein the port is in fluid communication with the second subchamber through the first opening to allow a stream of substance to be introduced into the chamber through the port substantially along a first direction.

A bioreactor comprising:

• a. a first substrate having a first surface, an opposite second surface and edges;
• b. a second substrate having a first surface and an opposite second surface, defining a cavity with a bottom surface, wherein the bottom surface is located therebetween the first surface and the second surface, and wherein the first surface of the first substrate is received by the second surface of the second substrate to cover the cavity so as to form a chamber for receiving cells and a liquid medium;
• c. a first filter dividing the chamber into a first subchamber and a second subchamber, wherein the first filter has a porosity to allow the first subchamber and the second subchamber in fluid communication;
• d. perfusion means in fluid communication with at least one of the first subchamber and the second subchamber to allow diffusional exchange of nutrients and metabolic byproducts with the chamber;
• and
• e. a port formed between the bottom surface and the first surface of the second substrate with a first opening and an opposite, second opening, wherein the port is in fluid communication with the second subchamber through the first opening to allow a stream of substance to be introduced into the chamber through the port substantially along a first direction.

A method for preparing a layered perfusion system 900 for use in a bioreactor, wherein the bioreactor defines a chamber for receiving cells and liquid medium, comprises the steps of:

• a. arranging a silicon wafer 953, a silicon-nitride layer 952, and a coblock polymer layer 951 such that the silicon-nitride layer 952 is positioned between the silicon wafer 953 and the coblock polymer layer 951;
• b. etching a plurality of channels 904c in the silicon wafer 953;
• and
• c. etching a plurality of pores 903c through the silicon-nitride layer 952 to form a filter 903 such that the plurality of pores are in fluid communication with the plurality of channels 904c.