Wednesday, October 22, 2008

The more legs you have, healthier cells you'll get
The shining dendrites show that the cell are dividing

The cell has changed into multipolar morphology

The confluent melanocytes after day-12

This is the story about my cell. I used to work with melanocytes or ‘mel’ the other name given by me. At first glance, I don’t have any idea on how difficult to handle melanocyte. In early of 2007, I bought the first vial of normal human melanocytes from Cascade Biologics. I can’t describe about what I feel for the first time to handle this cell. Before play around with melanocyte, I used to handle 3T3-L1 supplied by our senior Ting. For the first timer, I quite easy to handle melanocyte based on some knowledge from journals and articles. But I did a big mistake on the first step to thaw the cells. I used to follow the general procedure that being adapted on cell thawing. I forgot, that my primary cell is different from other cell line that we have in the lab. We can’t centrifuge normal cells. We just add medium then let the cells grow about 72 hours without disturb. For my first batch, the first and second passages are growing as expected. The cells grow beautifully with dendrites attached to the flask. I observed the cells, and then I know the cells grow from the dendrites. For melanocytes, the more legs you have, the healthier cells you get. I also get to know, the healthy cells are shining like a star. Suddenly, I start to have problem with the cells. I have a lot of dots in the flask that I can’t figure out whether it is contaminated or it just the byproduct from the cells. But the medium get cloudy and that moment I don’t have any experience to handle problematic cells. I could not safe my cells and even to finish my standard growth curve. Then, with the generosity of CEPP, I bought the second vial of melanocyte. At this time, the scenario is different. I was given a dead cells vial to thaw. Luckily, I have got my replacement from the principal. For the third vial, my heart still pumping fast like I never do the thawing procedure before because my cell is very sensitive. Form the findings, normal human melanocytes has passage limitation. Adult melanocytes can grow until 5 to 6 passages meanwhile foreskin melanocytes can survive until 7 to 8 passages. For the third vial, I managed to analyze the melanocytes for melanin and tyrosinase assay. I used to handle the cells on my own routine based on my experience and observation. When I see strangers in my flasks, I will increase the percentage of penicillin/streptomycin up to 2%. I need to change medium every 24 hours. The melanocyte is very sensitive, so PBS is restricted to wash the medium or else you will loose the cells. I used to wash using the medium so it makes the maintenance costly. You always feel difference when working with melanocyte. The best part handling melanocytes, when I have to talk with them and cuddle like my own precious baby. It works, even though they can’t survive till the end. When handling melanocytes, you have to shut off the light because it is sensitive to light. Don’t be shocked, the pellet of melanocyte is black in colour meanwhile most of the other cell pellet are yellowish. The cell count also not so exciting, the most I can reach is almost the power of 6. Many studies have been done using melanocytes, but most of researchers are prefer to use melanoma instead of using melanocytes because of melanocytes limitation. Whatever it is, it is wonderful to work with melanocytes and it teaches me how to be patient and think fast how to overcome the strange situation.

Monday, October 13, 2008

Fig. 1: 100% confluent of 3T3-L1 preadipocytes

Fig. 2: Mature 3T3-L1 adipocytes

3T3-L1 cell is a cell line derived from disaggregated Swiss 3T3 mouse embryos. The cells are able to undergo adipogenesis to form mature adipocytes. Here, the preadipocytes were cultured with DMEM (Dulbecco’s modified Eagle’s medium) supplemented with 10% fetal calf serum (FCS). Once the cells were 100% confluent (Fig.1), the cells were incubated with additional 48 h before initiating differentiation. At this time, 3T3-L1 preadipocytes expressed very early markers of adipocyte differentiation.

Subsequently, the preadipocytes were induced with a defined adipogenic media to differentiate the cells. The media is DMEM with a combination of dexamethasone, isobutylmethylxanthine, insulin and 10% fetal bovine serum (FBS). 2 days after induction, the isobutylmethylxanthine and dexamethasone were removed but insulin was maintained in media for another 2 days. Thereafter, the media were changed in DMEM with 10% FBS for every second day. 3T3-L1 cells began to differentiate 1-2 days after the cells were put back in DMEM and 10% FBS. Between day 8-12 post-confluent, the cells became fully mature and these matured adipocytes will continue to store triglycerides. As shown in Fig. 2, the red area demonstrated the lipid droplets, stained with oil-red O.

Adipocytes play a central role in whole body energy homeostasis. Therefore, a lot of studies can be done by using these 3T3-L1 adipocytes such as morphological studies, glucose analysis, metabolome profiling, assessment of cell growth/proliferation, and assessment of cell viability and apoptosis.

Wednesday, October 8, 2008

My turn--Nor Azurah Binti Mat Akhir (Azura)

Part A - about me:

Name: Nor Azurah Binti Mat Akhir
Age: 24 years old
Qualifications: Bachelor Degree in Biology (Genetics), National University of Malaysia. Currently pursuing Master degree in Bioprocess Engineering, University of Technology Malaysia.
Job Title: Research Associate
Employer: Chemical Engineering Pilot Plant and has been with this company for 1 1/2 year.

what drives me in my research:
Being a researcher is something worth to strive for. Exploring new thing and expecting the unexpected result, are the reason for me to choose this path. I am curious and this curiosity really fits me into this area. What really excites me is the exploration that I'll gain after my research is complete and lead to another new unexpected exploration.

Postgraduate studies:
this research is very meaningful to me as the result can give our traditional Malaysian herb one step ahead in contributing towards curing dangerous disease as cancer. This is a kick start for me as I can diversify my knowledge and skills in applying to other traditional Malaysian herbs, or widen up the usage of this one specific herb.

Part B- My background, my postgraduate study and current job.

Choosing this area of research/Postgraduate study:I have been exposed to molecular work before and I'm glad that I'm involving again in pursuing my master studies. I know that cell culture is quite apart from identifying DNA bands, but when we have strong basic lab working skills in molecular area, I truly believe that it helps a lot. I am also identifying the potential of our Malaysian Traditional Herb (Mas Cotek). I believe that every single herb that blooms around here in Malaysia has its own therapeutic value and this has driven me to choose the area of my study, to prove scientifically about what traditional practitioners have claimed, 'herbs that can cure cancer'.

Starting postgraduate studies:
i have already planned to further study in master's degree after i finished my undergraduate. I know that if this is the way that i choose i should be serious and equip myself completely. I have graduated 2 years ago, gaining experience in CEPP for solid annum and go for a master degree. I'm thankful that I am on my track and hopefully i can have doctorates degree before 30 \^O^/.

My research:
The morphological and growth characterization of human ovarian carcinoma cell line treated with
Ficus deltoidea (mas cotek) extract.

The best bits:
I learned lots of thing and will learn more things in the future. learning is process of life, opportunities that should be grab, patience, dealing with people, being knowledgable and the best is i do what i like to do - explore.

My Advice:
be yourself, live life positively and always be happy. Grab all the opportunities, leave the entire bad thing behind but learn from it, stay forward and focus. this recipe should be enough to motivate my self and with this, i am always looking forward, enjoy life and love to learn something new.

this is my story.....adawiyah....

PART A: About me

Name: Rabi’atul Adawiyah binti Ahmad
Age: 25 years old
Qualifications: Bachelor of Applied Science (Honours) (Biotechnology), Universiti Sains Malaysia, 2005. Currently pursuing Master in Bioprocess Engineering, Universiti Teknologi Malaysia.
Job Title: Research Associate
Employer: Chemical Engineering Pilot Plant (CEPP), University Teknologi Malaysia
What drives me in my work/research: My interest in doing research and I hope with this opportunity, I can learn new things and share my knowledge to contribute new inventions.
3 things I can’t live without: My laptop, hand phone and watch
Postgraduate study: I want to achieve my targets to develop nutraceutical products with scientific proven and fulfill the standard and requirement.

PART B: My background, my postgraduate study and current job

Choosing this area of research/postgraduate study:

I am a person who always looking for new experiences and hopes that I will gain more knowledge by doing postgraduate study. I have considered my own interest to further in bioprocess engineering. In future, with my knowledge and experience, I would like to join pharmaceutical industry to produce variety health supplement from local resources. Synthetic drugs with significant side effects may lead to other health problems. As for that reason, I hope that health supplement from natural resources can be as an alternative to synthetic drugs.

Starting postgraduate studies:

I started my postgraduate studies after a short course in ‘Herbal Plantation and Processing Technology’ and 6 months of working experience as a research assistant at CEPP.

My research/postgraduate study:

My research study is “Study of antidiabetic activity on cinnamon extract”. The purpose of this study is to investigate and prove the efficacy and toxicity of the cinnamon extract and its potential as antidiabetic. It was quite tough field of study since I have to familiar myself with ‘CINNAMON EXTRACTION’, ‘COMPOUND ISOLATION’, ‘ANIMAL STUDY’, ‘COMPOUND DETECTION ‘ and …… since UTM is an engineering university, I face difficulties to do ANIMAL STUDY here…=(

My first challenge is to purify the bio-active constituent name cinnamtannin B1 (CB1) using column chromatography. After one semester focus on the purification to get CB1… unfortunately, I am not succeed to isolate the targeting compound (CB1) but I’ve isolated other compound, known as coumarin. Hehehe…that what research are…rarely you get what you want and accidently you get something else… Recently, I am doing in vitro study on 3T3-L1 adipocytes to identify the potential of glucose uptakes and lipid droplet of cinnamon extract from different source.

The best bits:

Handling cell is the best experience for me. You have to care it as your baby…=). I have learned to motivate myself, always think positive, gain new knowledge and built up my skills in doing research. I also have expanded my networking while doing postgraduate studies.

My work:

In vitro and in vivo studies of antidiabetic activity on cinnamon extract. The experiences and skills help me in my work. Perhaps this study will help me in the future.

Looking ahead/Life after postgraduate study:

I love research and I would like to explore more and built my career in this area. Hopefully, with my new findings can give benefit to people.

My advice:

Although the going gets rough, always believe that you are though enough to make it…..ishoni gambarimasho….=)

Thursday, September 25, 2008

its me...

Rozilawati binti Abdul Gaffar
26 years old
Bachelor of Chemical Engineering Bioprocess, Universiti Teknologi Malaysia, 2005.
Job Title: Research Associate
Employer: Chemical Engineering Pilot Plant, Universiti Teknologi Malaysia
Has been with this company for 2 years
What drives me in my work/research: I love knowledge; hence within my research capacity I can explore new things and excite the industry with my invention.
3 things I can’t live without: my mobile phone, my grayish car and fruits
Postgraduate study: I want to go further with my research, make it as high end cosmetic products with empirical scientific proven.

PART B: My background, my postgraduate study and current job

Choosing this area of research/postgraduate study:
I believe in power of knowledge. Choosing postgraduate is one part of knowledge exploration in deeply and doing cosmetic science engineering is to make people look beautiful in nature ways. I have considered my own interest to further in bioprocess engineering and strongly attracted by the facilities provided here. In future, I would like to have my own brand on the shelves and published my work for others benefit. I am hoping that I can change the justification of cosmetic especially from the local market to make it as high end products with guaranteed quality base on research and gives my consultation to peoples who need it.

My research/postgraduate study:
My research study is “Study of Anti-tyrosinase Activity on Cashew Leaves using Melanocytes”. This study is about to explore the potential of cashew leaves which is proved by other researcher contains highly antioxidant and maybe act as good anti-tyrosinase. The challenge of this study is to conduct the analysis using in vitro analysis which is using melanocytes. At the end of the study, the percentage of tyrosinase inhibition will be determined. I am eager to polish my research skills, interpreting and analysis data and the most to expand my knowledge and skills in tissue culture. Hopefully, my research work will be a bench mark or standard to identify the quality of lightening products.

The best bits:
The opportunity to meet people, to deal with different level of people, working under stress condition but not most of the time, developing my own network, gain new knowledge and the most learn every single skill in doing research.
Looking ahead/Life after postgraduate study:
I am planning to pursue doctorate after completed my master. I love research and management work, so that I am not planning to switch my job to another field. I want to build my reputation and being an established consultant in this area.

My advice:
Pursuing postgraduate studies is such good opportunities to learn new research skills. For who is doing master by research, this is the pathway to conquer the knowledge in our research area. Don’t be afraid for the challenge, just believe in your skills and capability.

Wednesday, September 24, 2008


PART A: About me

Name: Rohana Binti Abu
Age: 25 years old
Qualifications: Bachelor of Engineering (Chemical), Universiti Teknologi Malaysia
Job Title: Research Associates
Employer: Department of Chemical Engineering (Bioprocess), Universiti Teknologi Malaysia
What drives me in my work/research: My mom
3 things I can’t live without: My specs, my only car, my meow meow
Postgraduate study: Master of Engineering (Bioprocess)

PART B: My background, my postgraduate study and current job

Starting postgraduate studies:
My research project entitled ‘The effects of free fatty acids on insulin receptor sensitivity and glucose transport in 3T3-L1 adipocytes`. Well it was quite tricky when I first read about my project. Coz, frankly speaking, I have no biological knowledge and this project is based on that knowledge. Erm… quite scary, isn’t it? But yeah nothing is impossible as long as you know exactly what you want to do. So, I take this challenge. So here’s the beginning of my stories….

My research/postgraduate study:
High level of plasma free fatty acid (FFA) is known to induce insulin resistance and that insulin resistance is linked to type 2 diabetes. The increased level of these free fatty acids is due to the increased consumption of fats which consequently reduced the sensitivity of insulin receptor and thus affected the glucose regulation in cells. Is this really happening? or it’s just propaganda behind the claims… nah! This is what I want to prove…like detective ah ha!

The best bits:
Handling the cell is the best thing I have ever done. Thanks a lot to my supervisor, Prof. Dr. Mohamad Roji Sarmidi (who first introduced me the project) and our head of research team, PM Dr. Fadzilah Adibah Abd. Majid for their kind supervision and continuous guidance and help. Thanks also to Ting Lee Yu, my senior, who taught me everything and my lab mates for their love and friendship (when I once nearly lost my ring!)

My work:
In vitro study is carried out to investigate the effects of free fatty acids on insulin receptor sensitivity and glucose transport in 3T3-L1 adipocytes.

Looking ahead/Life after postgraduate study:

The animal cell culture certainly suited my interest. So, I want to explore more cell-based work. Besides, I’m looking for great efforts to cure all diseases that killing people especially breast cancer as far as I concerned and as far as I could. With that in mind, I would like to pursue a PhD in the related field. After that I want to move on to do an academic postdoctoral position and continue within the academic community with the idea of my own research.

My advice:
When you do something, do it with genuine heart. Then you’ll be happy.

Monday, September 22, 2008

Human Skin Fibroblast 1184- husna

Picture (R)- HSF 1184 after 1 day revival
Picture (L)- HSF 1184 at 100% confluent
I use human skin fibroblast 1184 in order to achieve my research objective to determine the
cytotoxicity of papain-based wound cleanser to the healthy cells around the wounded area. HSF 1184 will also be used in order to investigate the cleanser’s effectiveness in promoting wound regeneration based on in vitro wound assay.

Human skin fibroblast 1184 was purchased from European Collection of Cell Culture (ECACC). It was derived from human skin by none other than Dr C. Arlett from the MRC Cell Mutation Unit, University of Sussex.

I’ve handled HSF 1184 since 21st August 2008. Picture attached here shows the cells after 1 day revival. For the first revival, it took about 7 to 8 days for the cells to become 70-80% confluent. It was tricky at first since I wasn’t sure about the cells’ pattern. It took me a while to get used to the cells thus the experience was indeed priceless.