Pushing Myself Back on Track

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.