Post on 27-Jun-2015
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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
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
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
Mathematical Model defining to Experimental Design
RSMIndependent variables:Collagen matrixSugar 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
EXPERİMENTAL DESİGN
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 variablesCell growing
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
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