Parameters of bioreactor, conditions to monitor

Flow rate of reactants/ growth factors

Concentration of product
Temperature
Oxygen flow rate
(the rest all in slides)
how much ECM molecules deposited (on scaffold)?
Different production methods:
Batch
Fed-batch
Continuous
How do the product look like from each technique
Simple prepare reactor sterilize close and let it run for few days,
stop culture and extract product
Start at defined point time, add glucouse with time, products
accumulate with time
Batch no medium supplied, shorter run time, nutrients run out and
therefore limited output
Continuous means fresh nutrients fed in while output is extracted,
like conveyer belt
Bioreactors also can mean water filtration systems? Why is bio
prefix there?
But not covered in this module, the classical applications
What requirements does a bioreactor need for tissue engineering?
pH, temperature, sterile easy to clean and maintain fits inside a
cell culture system, blah blah in front of slides, culture medium
monitoring, tissue engineering always has a scaffold, so handability
of scaffold and fixating it must be easy
good that if can apply mech. Stimulation to scaffold
adding growth factors to culture
3 main domains of bioreactor application
in slides
1. Pragmatic tool
2. Manufacturing device implants or grafts in clinical use
3. Modelling system
Pipette cell on scaffold is called cell seeding
Non repeatable or systematic manner of seeding
Therefore use machinery, cell seeding on 3D scaffold with optical
sensor and chamber with scaffold

Oscillating perfusion of cell seeding

Viable cells are significantly different, because
Reason why viable cells more proliferate in perfusion method of cell
seeding
1. With dynamic flow they get access to nutrients
2. Can cells die during process of cell-seeding?
a. If you just use pipette droping dead cells remain on
scaffold
b. But if dynamic seeding, dead cells drop off and not
adhere to scaffold
How to optimize dynamic seeding efficiency?
Cannot change scaffold composition, therefore have to look towards
cells
1. Flow rate of cell suspension that goes through the scaffold
2. Concentration of cell suspension, higher is better
3. Dont change scaffold porosity
4. Time period and injection pattern of perfusion, continuous or
interval (fast with pauses? Or burst and settle?)
5. Just drop it in, only live cells on surface of scaffold, in petri
dish, nutrient supply to cells is diffusion limited
6. In a mixer, have to consider how to remove waste products of
cells else environment becomes toxic
Point of physical conditioning
Mechanical conditions applied to scaffold does help tissue achieve
mech properties of fresh tendon
Therefore bioreactor must be able to allow experimenter to control
mechanical conditions of tissue-scaffold culture
Theres a summary slide for this
1. More efficient and homogenous seeding
2. Overcoming diffusional transport limitations
3. Increase biomechanical properties of engineered tissues
Another application of bioreactors modeling systems
Study how cells behave, whether vascularization is effective
Check compatibility of new scaffold material, implants or
grafts in (physiological conditions), multi-cell inter reactions in
physiological conditions, mimic disease conditions to test
cures, check whether nasal chrondrocytes behave as if knee
chondrocytes from knee cartilage expression of marker
proteins and check their differention
Good if we can harvest nasal chrondrocytes amplify and
implant back to repair knee
Model system slide for more study applications
Scaffold growing in reproducible and sterile conditions, allows
for scaling

Autologous limits the scalability because cells from patient 1 goes

back only to patient 1, i.e. many small bioreactors for cell-scaffold
culture is required for each patient
For each cell culture type, intricate details of that cell need to be
understood > requires different types of bioreactors for a particular
type of cell
Besides dictating flow rate of pump, we also have to check that flow
rate does indeed affect diameter of urethra diameter
Smooth muscle alpha actin is a marker for such protein presence
Double check PCR
Dynamic culture is better than static culture
Mechanical stable, tissue cultured from bioreactor
Breaks into groups of 2 or 3 to understand how commercial
bioreactors work