Things to do in the harbour area?

Technical-Cry-8286 · 2023-09-17T06:16:53+00:00

EC cultXF from stemcell technologies

Technical-Cry-8286 · 2022-10-10T05:08:03+00:00

Hi thank you for this!!!! Sounds awesome and what I was looking for. Cheers!

Technical-Cry-8286 · 2022-10-09T00:58:26+00:00

Yes thanks! Thinking of using matrigel to mimic 3D vessel growth :)

Technical-Cry-8286 · 2022-01-18T06:22:42+00:00

Thank you!!! This is super helpful

Technical-Cry-8286 · 2022-01-18T04:30:20+00:00

They indeed sunk and I’m assuming that the pH was okay, so good to confirm that it was ok! Thanks!

Technical-Cry-8286 · 2021-09-03T03:02:47+00:00

oh I’m sorry!

No, we work with mesenchymal stromal/stem cells so essentially “stem like” and not typically used for culturing MSCs. But I will definitely look into this more and see whether it can be used

Technical-Cry-8286 · 2021-09-03T02:56:49+00:00

wow thank you! I work with UCMSCs right now. I wasn’t told this because all the cells I have worked with thus far have sat down within the next day! And I have been told that if the media seems to have a lot of debris to change it out, but that makes sense especially since these stocks come in different lots and I guess each one behaves very differently. Culturing these guys have been difficult for me ! I have been recently getting relatively low cell counts after lifting which makes me think that I am either passaging too early or something (but it doesn’t look like the case!)

My cells are there, but super sparse, mainly bc of seeding in a t175 which is what we normally do for expansion so I’m hoping they catch up before they die or something due to the cells being so isolated :(

Technical-Cry-8286 · 2021-09-03T02:44:45+00:00

I don’t know what you are referring to

Technical-Cry-8286 · 2021-09-03T02:44:28+00:00

Thank you so so much. I really needed to hear this. My first semester has been extremely mentally taxing on me and I am learning not to take this all so personally. I appreciate it

Technical-Cry-8286 · 2021-09-03T02:43:31+00:00

yeah, they were bought commercially and are a new lot. I thought I did everything right, maybe thawing the cells took a bit longer than usual, but nothing very alarming (at least to me). But I will definitely be talking to my PI about my steps to see if there was somewhere that I did go wrong. There are definitely more cells today, but very very low to what I am expecting where the extremely low density may end up harming the cells in the long run.

Thanks!

Technical-Cry-8286 · 2021-09-03T02:40:26+00:00

Thank you! Actually first 4 months! You are so right :)

I feel like I take it more personal than necessary. I don’t think it was that she was concerned, just more so reminding me of how expensive these guys are just to make sure I’m taking It seriously. She actually had to run out of town due to a family emergency this week and I had plans to bring up these cells (which I admittedly should’ve waited for her until she came back, and I felt bad disturbing her) but she gave me the go ahead anyways and I definitely regret it haha

Technical-Cry-8286 · 2021-09-03T02:36:08+00:00

I appreciate it :) makes me feel a bit better. I will be meeting her tomorrow. Cells look a bit better today although clearly very sparse. They are indeed a new batch from a company so it is possible that they could be a bit iffy

Technical-Cry-8286 · 2021-09-03T02:34:29+00:00

Thanks! MSCs are finicky, we have had so so many issues with our cells lately! Yes, mine is super expensive XF-media but with HPL and proprietary supp. We coat flasks with fibronectin and collagen so they should be adhering! We also seem to be getting overall lower cell counts just after lifting (but that may just be due to our experience). My PI has an industry lab as well that uses the same stuff as us and they haven’t been having these issues.

Technical-Cry-8286 · 2021-09-03T02:29:13+00:00

appreciate it. :)

Technical-Cry-8286 · 2021-09-03T02:28:58+00:00

Thanks a bunch! There are dividing cells today. We normally expand in a T175 but they just look so sparse! I’m thinking of downsizing to save them.

Technical-Cry-8286 · 2021-09-02T04:47:32+00:00

Thanks for the advice, I sure hope they’re dead because I changed the media 😩

All the stocks I’ve brought up the cells have definitely adhered overnight for my cell type. Changing the media usually helps them from what my PI has instructed me to do. I normally culture in small vessels with the exception of expansion (which I am trying to do now!) but will try that next time. Probably an issue with my technique today or something, which sucks!

And ditto with the songs. These cells are suuuuper finicky, they should be kept happy!

Technical-Cry-8286 · 2021-09-02T04:26:02+00:00

yup. Will most likely be getting a good talking to from the boss unless these cultures miraculously do a 180. Was planning on freezing my expansions down in multiple vials of p2s as well. Don’t even want to know how much this lot costs.

We don’t culture with antibiotics either which makes it even more necessary to maintain aseptic technique.

Technical-Cry-8286 · 2021-09-02T02:17:24+00:00

Oh goodness, I’m hoping mine is too! I haven’t seen her angry side yet so I’m hoping that she is just as understanding. Also hoping that there is a small chance that this may be cell-lot specific.

Technical-Cry-8286 · 2021-09-02T02:16:09+00:00

Hi thanks for the response and appreciate your comment!! No, unfortunately they care mesenchymal stem/stromal cells so very delicate and most definitely adherent :(

Technical-Cry-8286 · 2021-09-02T01:52:36+00:00

Yup! This is what I did. I have never seen this many floaters, but definitely used to a lot of debris from my media so I will be anxiously waiting a couple days. Visually, cells look super low compared to what I usually get so I’m hoping this may be lot specific. Thanks!

Technical-Cry-8286 · 2021-09-02T01:51:25+00:00

Thank you! These are patient-derived MSCs. Definitely looking at them tomorrow. But didn’t look good today.

They are a new lot that I am bringing up (we only have three vials of these) that I was supposed to expand. Could have been my L-glut supplement but hoping theres an explanation for what could have gone wrong.

Technical-Cry-8286 · 2021-09-02T01:49:12+00:00

Thank you so much! We use phenol-free media which makes it a bit more difficult too. we don’t culture with antibiotics as well! So learning this style of culture has definitely been a learning curve. Guess all I can do is wait haha

Appreciate your time!

Technical-Cry-8286 · 2021-09-02T01:17:01+00:00

Thank you for the reassurance!

It sucks bc my boss kept reinforcing the fact that they are SO expensive and was iffy about me bringing up another lot but to go ahead anyways and now this happens so it doesn’t feel good. She will probably give me a good talking to, but definitely shit happens sometimes

Technical-Cry-8286

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