Fraser Leslie

Bioreactor- The bioreactor comprised an inner and outer housing, which were configured as coaxially
disposed, nested cylinders. The inner housing was designed as a rigid basket from polycarbonate in
which the scaffolds were kept press-fit during cultivation. The basket had a perforated lid and a perforated
bottom and was placed in the medium flow path for axial flow through the basket. The bioreactor system
comprised a bioreactor, a sterile fluid pathway (made of -sterilized PVC tubing, which had low gas
permeability) that includes a medium supply vessel, a pump, an oxygenator and a waste vessel.

Culture chamber
Reservoir chamber
Medium chamber
Gas and medium exchange system
Instrumentation for monitoring and control

A Hollow Fiber perfusion Bioreactor (HFPB) is characterized as a high density,

continuous perfusion culture system. The system consists of a medium reservoir,
waste reservoir, gas exchangers, a perfusion chamber and a pump. The system
consists of a set of thousands of semipermeable hollow fibers within a tubular
housing. The housing is then fitted to inlet and outlet ports. These ports can control
the amount of medium that passes into the chamber that houses the hollow fibers.
The ports also allow for any liquid that enters to pass through the interior of the
hollow fibers. These ports are connected to the peristaltic pump through the
perfusion tubing. The perfusion chamber is closed with a lid that has four slits which
the other end of the perfusion tubing is connecting creating a loop. The culture is
pumped through the lumen of the hollow fibers, allowing nutrients and metabolic
products to diffuse both ways across the fiber walls. The medium can either be
oxygenated and sent back the other way or an entirely new medium will flow
through.
The medium is kept in a separate chamber from the perfusion chamber
and holds all of the fluid that carries all of the cells, growth factors and proteins. The
general make up of a medium will contain Fetal Bovine Serum (FBS), Eagle Minimal
Essential Medium (EMEM), basic fibroblast growth factor, Penicillin-Streptomycin and
non-essential amino acids. The goal of the medium is to create an environment that
closely relates to the body which allows the cells to differentiate and grow. The Fetal
Bovine Serum is blood that has had all of the red blood cells removed and due to its
low level of antibodies and containing many growth factors common with eukaryotic
cells. FBS is common place with tissue engineering mediums as it is remarkably
close to human serum. Penicillin-Streptomycin is used to allow stop any sort of
infections that might occur within the cells and culture. It is important to protect the
cells from bacterial infection because it could ruin the entire kidney or tissue. Eagle
Minimal Essential Medium is the base of the mixture which is used to maintain cells
in tissue culture. EMEM contains salts, amino acids, glucose and vitamins, all things
necessary for cell maintenance. In order for the cells once they stick to the scaffold
need growth factors to activate their growth. The basic fibroblast growth factor that
is located within the medium is what will incite the production of an extracellular
matrix, or ECM which is pivotal in the creation of a tissue. The cells that will be
seeded and grow on the scaffold are also located within the medium. The cells will
come from the patient if that option exists because it will ensure that the kidney is
not rejected by the body because it does not recognize the cells. There will need to
be a few different kinds of cells as the kidney is not composed of just one type of
cell, which is why the Mesenchymal stem cells will be the most common cell
because they can differentiate into a plethora of cell types. Renal cells and epithelial
cells will also be employed to allow all parts of the kidney including the vascular
system.
For the gas exchange system within the bioreactor, it will consist of an
oxygenator, whose sole purpose is to add the right amount of O2 concentration

back into the medium. The bioreactor will be equipped with a microcomputer that
will monitor the temperature, pH level and O2 concentration of the medium. The
microcomputer will also possess the ability to maintain these levels on its own by
making sure the temperature stays at 37*C and the pH at 7. The pump will be
peristaltic pump which closely models how the body also pumps fluids. The pump
works by sending the medium out in chunks that are determined by the settings on
the pump. There will also be a waste chamber that will be used to collect the liquid
used after cell seeding as it is no longer useful.
Mass transport is needed as it is the way that the cells nutrients to be
delivered from bulk medium to the cells and waste products to be removed. The
transport of oxygen and nutrients from the medium to the cells will occur via
diffusion during cell seeding and by a combination of diffusion and convection
during proliferation or cultivation.