Bart Krist

Adam Mickiewicz University in Poznań | UAM · Developmental Epigenetics

Dear fellow scientists,

For our research in skin development we are collecting mouse embryos at different time points during embryogenesis. We now got an idea to do some further analysis for which it would be good if we would not have to use whole embryos, but only the epidermal layer. The later stages of embryogenesis we can dissect without too much problem, but earlier stages give me great difficulty. Therefore, I would like to ask if anyone has a good working protocol for epidermis isolation from E11,5 (or around this stage) mouse embryos?

Thank you in advance!

Hello everyone,

We are currently in the proces of ordering a new cel cluture incubator for our brain organoid cel culture. Now I never really had any preference for a specific type of incubator, but perhaps it is good to think about this before simply purchasing 'the cheapest option'....

So, which incubator is in your opinion the best for growing neurons, spheres and organoids? and why?

For our research on Huntington disease, we use a murine model with human muHTT and wtHTT on a Hdh-/- background. These ice unfortunately do not breed well, with only three litters in our breeding pairs in the last year...

Do you maybe have any tips for improving breeding capacity? We already changed from breeding pairs (1 male and 1 female) to a 'harem' system, with 3-4 females for 1 male.

All ideas are welcome.

Dear all,

I am trying to optimize siRNA transfection of primary sorted satellite cells from mice. Previously I had a lot of succes with transfecting murine myoblast cell lines with JetPrime (Polyplus), and now we want to bring our research into the primary isolated cells. However, collegues of mine have had very poor results transfecting SCs with JetPrime, Lipofectamine, siPort Neofx, and PEI  - low efficiency or high toxicity made succesful transfection previously impossible.

Does anyone have any experience with successful primary sattelite cell transfection?  which protocol you used?  Do you have any suggestions for further optimization? 

Thank you all in advance!

Hi everyone,

I am operating mice to induce ischemia in the left hindlimb by ligating the femoral artery. The procedure is well established i our laboratory and after three years of working on the project I might say we got quite experienced with it.

However, recently we see that mice of one certain genotype develop swelling of the right (=not operated) hindlimb. Infection is not likely, as the rest of the mice (operate leg, eyes, etc) are not inflammated, red, or swollen...

Does anyone has an idea what might be the reason of this swelling?

I would like to treat HUVEC cells with conditioned medium from transfected C2C12 cells. I was just wondering if C2C12 cells would survive if I would put them in MCDB-131 medium (which our lab uses to culture HUVEC) for 24hrs... does anyone has experience with that? or would it be more safe to condition DMEM medium and then dillute this in MCDB? - I would love to hear your thoughts and tips on this

I am planning to perform a global analysis of gene expression for the first time. I will be focusing on angiogenic factors. We are using mouse samples. Does anyone have any experience with such analysis? What are your tips and recommendations? PCR array, macroarray or microarray and from which company?

In our lab we use Ottix Plus and Ottix shaper dehydration and deparrafination media, as more safe substitutes of Ethanol and Xylene.

However, when I was fixing and staining some muscle samples, I saw the morphology was quite bad. Can this be because of ottix, or is my fixation (10% formalin solution for 48h) the cause?