Anyone have a cell lab yet?
  • CassoxCassox November 2015

    Just curious if anyone is farther along than I am in this forum regarding a biolab setup. I still have a ways to go. Lets say, theoretically, I had a nice pristine lump of stem cells. Does anyone have the resources to culture and utilize them? Theoretically of course.

    I wonder what the legalities regarding something like that would be? If some girl donated her aborted cell lump.. is this illegal to play with? What would you do with it anyhow?

    Thoughts?

  • bciuserbciuser November 2015
    So I have the general know-how and cell culture lab experience, but no, not the resources. Monies are a thing. As for the bare minimum for working with stem cells you'll need for the actual lab setup, though, here are the things you will need, from most to least expensive:

    1. A laminar flow hood (not a fully enclosed hood or a fume hood, but a HEPA-filtered positive pressure hood)

    2. An incubator capable of CO2 regulation (to be kept at 5% CO2 and 37C), and CO2 tanks.

    3. A 4C fridge for storage of recently made solutions

    4. A -20C freezer for short-term use aliquots (if you want to go crazy for REALLY long term storage, get a -150C as well)

    5. A lot of ethanol, and a spray bottle for the ethanol

    6. A lot of nitrile gloves

    7. Plates for culturing cells (typically we use disposable plastic, although you could probably use glass and sterilize between uses).

    8. An autoclave for sterilization (ideally one that can handle both liquids and solids

    9. A centrifuge capable of at least 1,100RPM

    10. Micropipettes of various sizes along with disposable micropipette tips

    11. A hemocytometer

    12. Some beakers and flasks for shits and giggles

    13. An appropriate labcoat (one that covers all your skin up to underneath where your nitrile gloves will cover).

    14. Conical tubes for use with the centrifuge.

    15. Larger tubes for media and solution storage

    16. A bunch of plastic junk to hold everything

    I may have missed a couple things and I'll add them back in if I notice. After that, the most expensive things to buy are the growth factors to keep the cells alive and multiplying, these vary significantly depending on what you are trying to grow (in fact, some stem cell culture media doesn't require growth factors at all). You'll also need to purchase the base media (typically some variant of MEM, with a shit ton of amino acids, salts, and other yummy stuff) and kits for cell dissociation (NeuroCult).

    When and if anyone chooses to move forward with the setup of a lab I can provide protocols and tutorials for most of basic stem cell culturing techniques.
  • glimsglims November 2015
    A lot of it comes down to that -150, just cause you want to be able to split and save you samples, not just do it once and then re supply each time.
    The -150 is basically a hefty dewar flask with liquid nitrogen in it. I believe that this is the biggest hurdle in getting set up, in as much as you really need to have a regular supply of liquid nitrogen.  Has anyone looked into building one of those nitrogen generators lately?

    The co2 is a pain, but just something you need to buy. Legally, you need a specific storage area for your tanks as well. 

    Also, before anyone gets google happy with the mushroom/orchid hoods, an addendum for the hood it that it is a vertical laminar flow hood, not a horizontal one. I'm working on a model with some of the people down in austin. Small, one person, relatively inexpensive. The plan is that once I'm done, I email them the plans and then they try to build their own. If it works, I'll post it up here so y'all can have one too.  There's a guy south of Seattle that will test the hood to make sure the pressure is legit.

    You can make your own cell dissociation mix with trypsin and bovine serum aaaand i think there is something else but I'm flaking. That reminds me though, you will want a vacuum pump to remove you media before you split your cells


    As far as legality, as long as you and the space pass all the biosafety 2 requirements, you should be fine. You have blood handling permit and a hep vaccination I would assume. Also, technically, you would register your "donated sample".

    I can also help out with protocols if anyone gets up and running. Last I heard, the Austin biohacker lab was getting really excited about organoids, and so they are becoming focused on getting a culture lab up...
  • bciuserbciuser November 2015
    And Pasteur pipettes for aspiration of said media. Just to clarify, a -150 is not the same as a liquid nitrogen freezer, which is closer to -195. Frozen samples should be stored in the nitrogen freezer, not a -150. Growth factors and antibodies can be stored long-term in a -150.
  • glimsglims November 2015
    Good point. My bad. I don't think I've ever needed a 150 before. We stored our stuff in a fridge or an neg80. This may just be a protocol difference...
  • JohnDoeJohnDoe November 2015
    Whould I need a cell lab to do research like this? I think that might be more practical than implanting a computer chip, at bare minimum it would have interesting implications.
    LINK:
    http://www.popsci.com/technology/article/2013-04/stanford-researchers-build-biological-transistor-within-living-cell
  • CassoxCassox November 2015
    Useful list. I compiled a damn near identical want list. The real question I'm getting at here is.. are there any interesting projects people can think of using human stem cells?
  • JohnDoeJohnDoe November 2015
    A bio computer is one, rapid muscle and tissue repair, maybe some sort of enhancement of the brain could be another. (Like a second brain by your gut talk about a gut feeling) more to come....
  • TheGreyKnightTheGreyKnight November 2015
    Well, the biggest target for me, if I were even going near HSC research, would be finding a way of creating usable stem-cell samples artificially without having to rely on new tissue samples. Renewability is really what I'm wondering about here. 

    I'd be interested in seeing if you can program them to grow custom sensory organs. Like, small nodes scattered around the body that are inherently sensitive to magnetism. Or maybe skin that glows when you walk into a dark room. 

    Another big spot for potential would be doing something like mitochondria 2.0, and adding a new organelle. 

    And finally, will stem cells trigger viable nerve interface growth between something with a slice of nerve tissue attached, and a severed nerve in the body. 

    Also, finding how susceptible they are to going into cancer mode would be nice. 
  • JohnDoeJohnDoe November 2015
    Do you think I could make a practical lab that could fit in a 9 cubic foot cube?(baring sample storage.)
  • bciuserbciuser November 2015
    @JohnDoe No. Maybe 9 cubic meters, if you're REALLY good at Jenga. And 4 feet tall. And have tiny hands.

  • bciuserbciuser November 2015
    @TheGreyKnight

    Full tissue culture that can be integrated into the human body (that actually does something, I'm not talking about tracheas here) is quite a ways away, and not something that can realistically be done at this point.
  • glimsglims November 2015
    Yeah, sadly, some of these things take space, even without the fridges. I think I have a picture of working in a space like that....

    http://imgur.com/g0I1yVg

    aww yeah, look at that . The chemE dept didn't like giving up space to the bio people...
    So that's cramped ass lab right there. It's about 9x9x12 . But you can see how there is a emergency exit door all weird in the back corner and a second fridge. You could scrap one of those fridges if you're working by yourself. The incubator is out of shot but could go on the left of the hood. Sink is on the right.
    I bet you could get this to fit in a standard shipping container, with a lot of good planning...

  • JohnDoeJohnDoe November 2015
    Okay never mind I get that it was a bad question.... I have plans to start wondering soon and that was why I asked.
  • bciuserbciuser November 2015
    Not a bad question, as I assume you have no cell lab experience you have no way of knowing the size requirements of a lab.
  • JohnDoeJohnDoe November 2015
    Yea I have never set foot in a cell lab before. Didn't think that it was even possible to build one till I saw this thread. I still want to I just got to look harder than what I am. It seems you have a better understanding however. May I ask you what the standard size of a cell lab is? I also would like to state that size constraint was for storage, it can fold out in anyway.

    Sincerely,
    John Doe

    P.S. For what its worth I am looking at making a super allege to power a van of of biofuel.
  • you can prevent cell from damage by ultra low freezer work on low temperature principle on 80 to 85 c. you are going to start new lab so people are scare to come to your lab because of you don't have experience for this but if you provide all facility to your patient then they will come to you again and if you are researcher then you have to prevent your sample from damage for your research result.

    http://newmeditech.com/cold-storage-equipment/ultra-low-freezer/