Hey all,
Brief context: I am looking into whether or not metal homeostasis plays a role in Salmonella pathogenesis and response to oxidative stress. Neither myself or anyone in my lab has significant cell culture experience outside of the one type that we use for some simple assays; we're almost exclusively a prokaryotic lab, so I don't really have anyone I can easily go to for technical questions.
Currently, my lab uses J774.1 murine macrophages for our intracellular Salmonella replication/infection assays, but after doing some reading I realized that J774.1s are NRAMP1-/- (metal ion transporter) and sought out a macrophage cell line that was NRAMP1+; which is how I learned about the HeNC2 cell line. From what I understand you cannot buy these commercially, so I cold-emailed a lab who seems to have given other labs some of their stock and asked if they wouldn't mind sending us some too and they sent us a vial (yay!). I tried to double-check with that lab that the media we use for our J774.1s would also work for HeNC2s, but never heard back (not yay), so my PI encouraged me to just start them anyways.
I go ahead and start them with the same media (RPMI-1640 + 10% FBS + 2mM L-glutamine + 15mM Hepes Buffer + 1mM Sodium Pyruvate) and protocol we use to start our J774.1s. I started them a little less than a week ago and since then they haven't really attached to the flask at all (or only a few have attached, but haven't grown. It's difficult to tell) and appear to be just floating in the media and not growing. The vial that was sent was from 2015 and the lab had mentioned that they're old but "should still be good". From what I can glean from the literature, they absolutely should be an adherent cell line similar to J774s, so I'm wondering if I did something wrong and killed them, or if I just need to be patient and wait longer for them to "wake up" since they came from an old vial. I've also been successfully culturing J774.1s (mostly successfully, I hate how they can clump sometimes if I'm not careful, but that is independent of the media) in the same media I used for HeNC2s, so I don't think the media was contaminated?
Anyways, I can provide more details on exactly what I did if needed, but I would much appreciate any thoughts or pointers from someone has worked with this cell line before or has general experience with immortalized macrophage cell lines.
Here is a link from Cellosaurus with a
... keep reading on reddit ➡Henrietta Lacks was an African-American woman whose cancer cells are the source of the HeLa cell line, the first immortalized human cell line and one of the most important cell lines in medical research. An immortalized cell line reproduces indefinitely under specific conditions, and the HeLa cell line continues to be a source of invaluable medical data to the present day.
https://en.m.wikipedia.org/wiki/Henrietta_Lacks
I was wondering if anyone here has experience with HIV and/or B cell lines. I’m designing a hypothetical experiment and I’d like to focus on non T cells but the research with HIV on non T cells is lacking or perhaps I’m just not reading it enough to understand it.
The non T cell papers I see are studies using primary cells and I would prefer to use immortalized cell lines if possible for feasibility purposes. The only cell culture experience I have is with 293T and MT2 cells and they’re pretty common in the field. I’ve never had to pick which cell line to use myself. How would I even go about finding immortalized B cell lines?
Does anyone here use these types of cell lines? Does anybody have a resource on where I can look at different cell lines? All I’ve been seeing when I try to look for it is ordering them but they don’t give a good explanation of the differences between the cells or what they’re commonly used for and I lack the background knowledge to be able to figure that out for myself.
Since immortalized cell lines are being used in common differential expression protocols how is it we take into account the technical noise produced by the cells themselves?
Hi everyone!
Does anyone know of an immortalized b-cell line (preferably with the cd20+ marker) that can be used in a biosafety level 1 lab? If so, can you provide a link to where I can purchase it from? (I'd prefer to work with a human cell line, but if you know of a cell line from another animal, please share!)
Thanks!
I understand that normal cells have an upper bound on the number of cell divisions, does the "immortalized" imply that this cell line does not experience the shortening of telomeres during cell division?
How do we create immortalized cells? Are they cancerous? Are there any side effect of using such cells in genetics?
http://www.sciencedirect.com/science/article/pii/S0963689798000402
Would be very much appreciated if anyone has access!
Hi everyone!
I'm trying to order immortalized cell lines for the first time (undergrad) for a study I'm conducting this summer. I need normal thyroid cells from either a mouse or rat. So far I've seen a lot of thyroid cells with carcinomas, but only one that's not diseased (from the ATCC). Anyone have some other recommendations? I'd like to bring my research advisor a couple options we could discuss.
Thanks!
http://pubs.rsc.org/en/content/articlelanding/2013/mt/c3mt00149k
http://www.ncbi.nlm.nih.gov/pubmed/?term=Regullation+of+intracellular+pH+by+immortalized+human+intra-hepatic+biliary+apithelial+cell+lines
For someone who really has a good grasp on the subject, could stem cells be used for reverse aging therapy? Could we take a senior and with the right diet and exercise, and exposure to stem cells, they could revert to the prime of their youth and reset the clock on their mortality?
If this is possible why aren't we more loud about making this legal and a thing?
I'm trying to sell all of these as sets because my collection is large and packing and shipping takes time. However, if a set doesn't sell within three days I will split.
Hellsing 1-3 (g5) $90
Blade of the Immortal 1-4 (G5) $100
Cells at Work 1-6+ 1 Black (g5) $50
Rent a Girlfriend 1-4 (g5) $35
Vineland Saga 1-11 (g5) $135
A game that is so laconic as to have zero dialogue leaves much in the way of room for interpretation. Rife with mystery and riddled with environmental clues, the storytelling method of Hyper Light Drifter is recondite and abstract. Piecing together the details that we come across, we can begin to construct a cohesive narrative of the concrete historical events and underlying factors that have contributed to the experience that is played through. By so doing, we stand to gain a deeper understanding of the metaphor that is conveyed through Hyper Light Drifter's story.
In a response to a previous thread I speculated that the Perfect Immortal Cell was initially crafted to refine a neon-pink substance that is widely present throughout the land of Hyper Light Drifter. I am here to elaborate on that conjecture. In this thread I will conduct an in-depth exploration of the various applications of the neon-pink substance that is ubiquitously observed throughout the game, hereafter referred to as Hyper Light, its relationship with the Perfect Immortal Cell, and the effects that its production may have had on the land's inhabitants.
General Observations
Perhaps the foundational question that we need to ask on this topic is which came first—Hyper Light, or the Perfect Immortal Cell? In order to answer this question, we must establish from where they both originate, to wit: the Great Wellspring.
Starting from the beginning of the opening cutscene, we see particles of magenta wafting into outer space as our gaze pans downward, coming to a rest on the sight of a gleaming megalopolis at night. Floating above the city and giving off a tranquil blue glow is a rather elaborate ethereal construct. Now this could be the Great Wellspring that the Librarian's record tells us about. After all, by the look of things it appears to be pouring down on the city from above, like a fountain of distilled essence.
But I actually think that the opposite is the case. This construct is not the Wellspring itself, but I postulate that this is instead some effect of the Wellspring being projected into the sky from the
... keep reading on reddit ➡Henrietta Lacks was an African-American woman whose cancer cells are the source of the HeLa cell line, the first immortalized human cell line and one of the most important cell lines in medical research. An immortalized cell line reproduces indefinitely under specific conditions, and the HeLa cell line continues to be a source of invaluable medical data to the present day.
https://en.m.wikipedia.org/wiki/Henrietta_Lacks
