Meeting Summary Page
Meeting Date: 10 July, 2009
Summary:
01- Micro Stem Cell Chip and Automated System for optimizing the differentiation of stem cell using mechanical stimuli
- Fabrication and visualization of stem cell chip
Action Plan: (meeting comment)
1) RNA isolation using QUIAGEN RNase micro kit
- Macro-sized exp.
- Micro-sized exp.
2) Q-PCR test for gene expression
>> Macro-sized exp. using T-25 flask
- 3germ layer test: Mesoderm specific test using mesoderm specific marker (Brachyury)
Brachyury of macro-stimualtion sample expressed 2.8 times higher than macro-control.
- Chondrogenesis test (specific markers: Aggrecan, Collagen type II)
No significant expression
- Osteogenesis test (specific markers: ALP, Collagen type I)
Collagen type I of macro-stimualtion sample expressed 2 times higher than macro-control.
>> Micro-sized exp. using stem cell chip
- 3germ layer test: Mesoderm specific test using mesoderm specific marker (Brachyury)
Brachyury of micro-control sample expressed.
- Chondrogenesis / osteogenesis test
Several genes expressed in micro-control sample. But, there is no gene expression in stimulation sample.
--> Gene expression of micro-control sample shows that RNA isolation method using RNase micro kit is successful and useful.
--> No gene expression of mciro-stimulation sample.
; Cell would be died (1psi may be too high) or contamination ==> Next experiment: cell viability test on 0, 3, 5, 7 days
Next Plan:
1) Fabrication devices for the next mechanical test
- 6 or 8 devices (Low, 1 psi, High condition)
2) EB encapsulation in gel (gelatin-MA 5%, 10% and PEG 10%)
- Compare natural and synthetic polymer encapsulation
- Investigation of EB differentiation (vasculogenesis) in the polymer encapsulation
- 36 samples
Meeting Date: 25 June, 2009
Summary:
01- Micro Stem Cell Chip and Automated System for optimizing the differentiation of stem cell using mechanical stimuli
- Fabrication and visualization of stem cell chip
Action Plan: (meeting comment)
1) Culture 7 days with mechanical stimulation (conditions are same in the former meeting summary)
- Macro-sized exp.: 2 flasks (1 x control without stimulation, 1x stimulation)
- Micro-sized exp.: 2 stem cell chip (1 x control without stimulation, 1x stimulation)
2) Collect samples after Trizol treatment
- Collecting volume: Macro-sized exp.: 1 mL /each sample, Micro-sized exp.: 80 uL /each sample
3) UV-curable geltin patterning and preliminary culture-test
- Patterning geltain structure in micro chambers of micro stem cell chip (succeeded) <-- Sandeep
- EB Cell viability test in gelatin
--> Interesting result: many EBs sprouted and vasculized in geltin
(CD31 staining --> sprouted structure was stained)
Next Plan:
1) RNA isolation using QUIAGEN RNase micro kit
- Macro-sized exp.
- Micro-sized exp.
2) Q-PCR test for gene expression
- 3germ layer test
- Chondrogenesis test
- Osteogenesis test
3) EB formation and culture for geltin encapsulation exp.
Meeting Date: 15 June, 2009
Summary:
01- Micro Stem Cell Chip and Automated System for optimizing the differentiation of stem cell using mechanical stimuli
- Fabrication and visualization of stem cell chip
Action Plan: (meeting comment)
1) Mechanical stimulation experiments
- Macro-sized exp.: T-25 flask (Pressure: 2 psi, Frequency: 1 Hz, During: 10 min, every 12 hours)
- Micro-sized exp.: Stem cell chip (Pressure: 1 psi, other conditions are same)
2) Analysis total cell number of various sized EB (embryoid body)
- Total cell number of one pair cell chamber: 8,840 cells (100 um - EB x 17 wells x 2 chambers)
- It is enough to do PCR --> consider high efficiency RNA isolation method
Next Plan:
1) RNA isolation and PCR test for 3 germ layers test
- Macro-sized exp.
- Micro-sized exp.
2) Fabrication devices for more mechanical test
Meeting Date: 19 March, 2009
Summary:
01- Micro Stem Cell Chip and Automated System for optimizing the differentiation of stem cell using mechanical stimuli
- Fabrication and visualization of stem cell chip
Action Plan: (meeting comment)
1) Additional short-term application
- Oxygen concentration application (easy exp. and fast result - cell death)
2) Consider cell number of each chamber and single cell PCR
- Minimum cell number of PCR would be over 10,000 cells.
Progress on issues from last meeting:
1) Survey & verify
- Literature survey about oxygen concentration cell exp.
2) Survey & verify
- Literature survey about single cell PCR and consider about integration
Meeting Date: Jan. 6, 2009
Summary:
01- Micro Stem Cell Chip and Automated System for optimizing the differentiation of stem cell using mechanical stimuli
- Design test chip for stem cell chip using mechanical stimuli
.Novelity: This device is an easy tool to compare the differentiation of stem cell under mechanical stimuli.
.Target cells: mES, MSC (mesenchymal stem cell), Adipo cell
02- Microfluidic System for Screening Stem Cell Microenvironments (Dr. WonGu Lee)
- Prepare mESCs for cell loading exp.
03- mES Cell Encapsulation (Dr. Bonggeun Chung)
- Prepare mESCs for cell loading exp.
Action Plan:
01- Micro Stem Cell Chip and Automated System for optimizing the differentiation of stem cell using mechanical stimuli
- Literature survey
- Make mask
02- Microfluidic System for Screening Stem Cell Microenvironments (Dr. WonGu Lee)
- Expand & stock mESCs
- Cell loading exp.
- Fluidic control & Mixing exp.
03- mES Cell Encapsulation (Dr. Bonggeun Chung)
- Fabricate PEG microwell devices x 48 pc (10% 1000, 30% 1000)
- Expand & stock mESCs
- PEG diffusion test for simulation (using 10KD, 20KD FITC)
- Cell seeding
Progress on issues from last meeting:
Ali's comments:
How do you know mechanical properties are going to have any effect on ES cells?
What is the title of your paper?
We should move faster.
We should have some actual devices for the next group meeting.
Figure out the biology and fabricate the device
Meeting Date:
Summary:
Action Plan:
Progress on issues from last meeting: