3. Patent Research
Coated fiber scaffold for three dimensional cell culture of
neural cells, this is the important experiment for my
hypothesis[1]
Three-dimensional cell and tissue culture system, in this
research, collagen is used for encapsulated for cell[2]
Neural stem cell-collagen-bioreactor system to construct a
functional embryonic brain-like tissue. These neural
progenitor cells in the bioreactor were held arrest collagen
matrix[3]
Apart from these neural stem cells and neural cells in three-
dimensional collagen matrix with a lot of encapsulated is
under patent protection.
[1] LIU, J., Pekny, M., Puschmann, T. B., Zanden, C., (2012), Coated fiber scaffold for three dimensional cell
culture of neural cells, publication number: WO2012158120 A1.
[2] Naughton, A. B., Naughton, K. G., (1989), Three-dimensional cell and tissue culture system, publication number:
WO1990002796 A1.
[3] Ma, W., (2009), Neural stem cell-collagen-bioreactor system to construct a functional embryonic brain-like tissue,
publication number: US7588938 B2
4. Which is the best way?
O'Connor and colleagues observed entrapment of cortical neurons with collagen matrix
showed that exhibit behavior closer to the truth in their experiment [4].
[4] O’Connor, S. M., Andreadis, J.D., Shaffer, K. M., Pancrasio, J.J. and Stenger, D.A., (1999) immobilisation of neural cells in
three-dimensional matrices for biosensor applications. Laboratory for Biosensors and Biomaterials, Center for Bio:Molecular
Science and Engineering, Na6al Research Laboratory, Code6910, 4555O6erlook A6enue SW, Washington, DC20375, USA.
• Entrapment on sugar
poly(acrylate).
•Conformational properties of
the cells can not fully protect
Entrapment on collagen matrix
5. WHY COLLAGEN
MATRIX?
have large pores
simple gel formation
have high mechanical
strength[5]
injectable forms are unable to
sustain
the mechanical and shape
features [6]
[5] O’Connor, S. M., Stenger, D. A., Shaffer, K. M., Ma, W.,
(2001), Survival and neurite outgrowth of rat cortical
neurons in three-dimensional agarose and collagen gel
matrices, Neuroscience Letters 304 (2001) 189-193.
Washington, D.C. 20375, USA
[6] Bellamkonda, R., Ranieri, J. P. Bouche, N. Aebischer, P.
(1995), Hydrogel-based three-dimensional matrix for
neural cells, Division of Surgical Research, Centre
Hospitalier Universitaire Vaudois, Lausanne University
Medical School, Switzerland.
WHY rotating wall
vessel reactor?
•Supports three-dimensional
environment. All forces balance each
other.
•Facilitate diffusion of nutrients.
•Matrix synthesis speeds.
•Redifferentiation and stimulates
proliferation.
•Properly mixing reduces the
voltage[7]
[7] http://www.mderl.org/demos/brew.php
6. Mathematical Model defining to Experimental Design
RSM
Independent variables:
Collagen matrix
Sugar poly(acrylate) hydrogel
RPM?
Temperature ( 4 oC is ideal for
my cells)
I can not use RSM technique for these
Design
Expert 7.0.0
Response: Cell activity
7.
8. Independent variables
•Collagen matrix
•Sugar poly(acrylate) hydrogel
•RPM
•Temperature ( 4 oC is ideal for
my cell)
•No control group.
•Results from samples taken and the effect of the parameters is according to the
microscopic observation.
•Data analysis will be based on microscopic observations and viable counts.
•Independent variables changing causes a negative effect on cell growth, cause in optimal
conditions cells exhibit growth.[7]
[8] Borkenhagen, M., Clemence, J.-F., Sigrist, H., Aebischer, P., (1998)Three-dimensional extracellular matrix engineering in the nervous system. J.
Biomed. Mater. Res. 40, 392 – 400
Operating Mode: Fed Batch
Dependent variables
Cell growing
9. Result
Done during the assay culture system
measurements, increase in the
number of cells per day are expected
to occur. This is estimated supported
with current research[9]
Samples of the cells to the cell due to-
day there is an increase in the
number.
•Cullen, D. K., Vukasinovic, J., Glezer, A. (2007),“Microfluidic Engineered High Cell Density Three-
Dimensional Neural Cultures,” J. Neural Eng., Vol. 4, No. 2, 2007, pp. 159–172.
In this experiment results due to:
•microscopic observation
•The increase in cell number and cell growth
