It’s been a while since I last blogged, and although I have
promised the last time to blog on my experience of doing my Masters in Dr.
Adler’s lab, I have failed to get myself writing on the post as I am dwelling
in how much time it would take for me to write. I know, another form of
procrastination. I have recently just watched a video on how to improve on the
skills you want to build or make, and all it takes is to work on it daily. Yes, you
got it... watched it on ‘Nas Daily’ where he makes 1-minute videos daily, and
it doesn’t matter how much engagement you get, all that matters is that the
content touches your core, it is what you want to produce. There are times that
because you are pushing to do it daily, your motivation starts to fade, or
maybe you might be producing something without quality… but as time progresses
you would become better and better at it, naturally. I guess, this is something
we all know, have known and will have to constantly remind ourselves… as the
saying goes ‘practice makes perfect’. I
was totally inspired by the video and got myself thinking, hey, I should really
stop making excuses. Even if it takes a couple of sentences a day, never stop
writing… never stop progressing. So this topic goes to my attempt of studying
Japanese and Spanish on duolingo. If you were to ask me what my weakness would
be back in school, it wouldn’t be math. I was never scared of math, not that I
am good at it… but math has always been a subject at school I have enjoyed
doing. My biggest arch-nemesis was learning languages. I have the weakest language
skills. Back in my school, we had to take Arabic as a foreign language. Despite
learning it for years and years, I was only able to learn so much. Yes, I am
able to read and write, maybe understand a few basic words, but that’s it. My
incompetency in picking up languages is basically obvious because despite
watching Japanese/Korean/Taiwanese shows for years and years, just
like some of my other friends, they are better at understanding and confident
at conversing than I am. So recently, I decided to challenge myself to learn a new
language properly, however slow I am at picking it up. Just so that I could
prove my own self, wrong (I have this thing that I like to do, I enjoy trying new
things --- so you could tell how many things there are on my bucket list! So much yet to cross off and I keep adding new to do's on my list). So coming
back to what I really wanted to emphasize, is the need of practicing the 2
languages that I have started learning daily, and with this practice… I am also
learning at becoming more consistent with my hobbies.
Okay,
enough of chitchatting. The major reason
I made this blog is so I could write on scientific findings, or knowledge that I
have gained either through attending a seminar, or reading a journal etc so that I get to understand things better, get the knowledge implanted in my head and at the same time share and spread the knowledge! So
for the first scientific knowledge post, I would like to report on what I have
learned from a seminar last Monday in UM. The seminar was on the ‘Innovative Strategies in Stem
Cell Research’. Dr. Roland Leather from Thermo Fisher Scientific was the
speaker for the seminar and of course, a talk given by a company would be biased
to promote their product. But, excluding the biases of the products used, it was very
informative and helpful for people like me who is still quite fresh and new in
the stem cell research line.
The first
talk was entitled the Innovative Solutions For Stem Cell and Neurobiology Research
Workflow. Dr. Roland started with introducing iPSCs and the workflow in
generating human pluripotent stem cells. As we know, induced pluripotent stem
cells are stem cells generated from our adult somatic cells and reprogrammed to
become pluripotent stem cell like cells. There are a lot of benefits and
potential in theory of iPSCs and one of the biggest positive factors of it is
that it is not immunogenic to its donor as it is from your own cell. Pluripotent
Stem Cells are highly preferred compared to mesencymal stem cells as it could
give rise to any types of cells from the mesoderm, endoderm, and ectoderm layer
of the embryoid body in comparison to mesenchymal stem cells that could only
give rise to a limited number of cells such as from the osteoblasts (bone cells), chondrocytes (cartilage cells), myocytes (muscle cells) and adipocytes (fat cells) lineages.
So the
workflow of creating induced pluripotent stem cells starts from taking adult
somatic cells from the patient and reprogram the cells by transfection of
specific transcription factors (ie. Sox2, Oct4, Klf4, C-Myc) that will
reprogram the adult somatic cells into induced pluripotent stem cells. Transfection
can be done using may different gene delivery technologies such as using
lentivirus, episomal vectors or the sendai virus. However, current issues with
the usage of lentivirus or retroviruses are the possibility of integrating
foreign DNA into the genome, which may lead to insertional mutagenesis. Hence,
using a non-integrating delivery system such as the episome or sendai virus, is
recommended. Episomes are circular extrachomosomal DNA molecules that carry
viral elements and can be transfected into the cells without the need of viral
packaging. Sendai virus on the other hand is a single-chin RNA virus and does
not integrate into the genome, hence no genomic footprints left and no
genotoxicity.
After
reprogramming, the iPSCs are cultured. Challenges that companies are facing
currently is to formulate culture media are that there are poor cell survival
following single cell dissociation, slow and variable recovery rate and
aberrant. There is much new advancement in stem cell media formulation, and one
of it tackles the issue of having to change media every day. There are also
many culture systems feeder or non-feeder dependent culture systems these days.
Following culture of iPSCs, the iPSCs can be characterized by TaqMan, alkaline
phosphatase assay, or the formation of teratoma and could after that, undergo
genome editing, either by CRISPR Cas9 technology or TALs.
After gene
editing, the iPSCs can finally be differentiated into different cell types of
cells. Induced pluripotent stem cells are like Embryonic Stem Cells and can be
differentiated into the cells of the three germ layers Endoderm, Ectoderm and
Mesoderm. The speaker got to touch on the differentiation into neural cells
only in this session. iPSCs can be differentiated into different neural
lineages, and in order for us to differentiate the iPSCs into neurons or
astrocytes, iPSCs have to be differentiated into Neural Stem Cells (NSCs)
first, then into neural progenitor cells and finally after that to neurons or
astrocytes. NSCs can be banked at day 16, which makes it convenient for the scientists,
as there is no need to reculture iPSCs. At this point, the speaker recommended
to use ‘culture one substrate’ as it helps NSCs to differentiate specifically
to neuron cells and avoid proliferation of NSCS so there is no interruption of
differentiation into neuronal cells. Finally, the diffentiated cells can be
characterized or measure by FACS, AP live stain, live cell staining kits, fixed
cell immunochemistry or Taqman hPSC scorecard.
I apologize for not getting into too detail, because honestly I got tired of writing 😂
I will continue the Part 2 (Thermo Fisher Capabilities on
Mesenchymal Stem Cell Application) and Part 3 (Thermo Fisher CAR-T Solutions)
of the talk on my next post! Promise xx.
Stay tuned.
AA.
Stay tuned.
AA.
