How can I grow a brain in a petri dish (seriously)?

A couple years ago (link), a fellow took some rat brain tissue, kept it alive in a solution in a petri dish, connected wires to it, and successfully used the brain to control a flight simulator.

I want to duplicate all or part of this experiment in my basement.

I can do all the stuff from the wires out, the wiring, amplification, signal analysis, software, etc. Well, I don’t know the exact formulas for everything, but I know how it’s done in depth and have confidence I can do it or find the right people to help me do it. But the rat brain living in the petri dish part, I have no idea where to start there.

Can anyone give me any pointers? I’m guessing these are some of the areas of concern:

  • Keeping the tissue alive. I’m supposing there are special fluids the tissue needs to be immersed in. What are they? Do I need to regularly refresh these fluids? Do I need to keep them oxygenated or anything?
  • Cleanliness. I’m supposing everything has to be super sterile from the beginning. Are there special tools or clean-rooms or anything that I need to work in while the petri dish is open, to ensure the internal environment stays clean?
  • Acquisition of rat brain tissue. I’m supposing you need to acquire it from a freshly killed rat, or even a live anesthetized (or non-anesthetized yet restrained, ouch) one. There must be techniques in this area. I’m not terribly squeamish, but I don’t want to kill any more furry friends than necessary.
  • Other. I’m certain there’s other stuff I haven’t thought about. Any thoughts will be appreciated.
  • Laws. Anything I need to worry about before undertaking something like this?

Thanks very much.

If it was that easy…!

I’m curious what you’re going to do with this?
Help with your taxes?
Home alarm system?
Cheese spoilage detector?

You’ll need Ringers solution. As you’ll only be dealling with neural tissue, you need to -

put nutrients in the fluid

remove waste products

oxygenate the fluid

remove CO2
etc.

This is not a goldfish in an aquarium. This isn’t even a disembodied organ. You have a type of specialized cell without any of the other organs or organ systems needed to keep it alive.

You need a clean room. This will require specific materials (some types of concrete, plastic, metal etc can be penetrated by outside pathogens, or are permeable to outside gasses). You need a humidifier, dehumidifier, an air conditioner, and a heater. All of these must be new and must themselves be kept carefully isolated from outside contamination. You need an airlock between the clean room and the rest of the world. Unless you have the greatest petri dish ever made, it does not especially matter if it’s open or closed.

I see no reason why the members of the genus Rattus available at pet stores everywhere would be unsuitable. But, you may need fetal or embryonic tissue. Anasthetic is likely illegal to obtain without a proper license (EG a veterinarian or the biology teachers at the local colleges has no problem buying ketamine. The general public cannot legally buy ketamine). The necessary surgical tools are probably legal. The type of needle used to obtain a neural biopsy could presumably be used to inject a human with illegal drugs, but it would be a waste of resources. A hypodermic for shooting up is thinner, less durable, and cheaper.
BTW-If you have reservations on killing rats, this is not the experiment for you. Even if it is possible for a professional with years of training to extract brain cells without doing any harm to the rat, you won’t be able to do so. Rat brains are not big things. A slip of milimeters can result in a rat that is dead or permanently brain damaged. You have not had years of study in sterilization of tools and operating rooms. You have not had years of study in the proper way to restrain a rat while taking a neural biopsy.

(Attempting to avoid a GD hijack- I support the killing of animals in the interest of saving human lives, or expanding the boundaries of human knowledge. I oppose the needless killing of animals. As you don’t seem to have the requisite degrees, or to have spent years studying the relavent fields, your work will almost certainly be unsuccessful.)

Electricity, heat, and water to your clean room must not be interrupted at any time. All systems for keeping the room sterile, at proper humidity and temperature, for providing nutrition and oxygen to the neural tissue, and for removing waste products must have multiple redundant backups. All electrical equipment in the room must have multiple redundant shielding to prevent electromagnetic interference with the neural tissue. All electrodes connected to the tissue must be medical grade.

You may or may not be in violation of animal cruelty laws in your country, state, or county. You may be in violation of zoning laws.

BTW

The clean room and airlock must be physically capable of withstanding intruders. You do not want a friend or relative to wander in for a look an contaminate the room. PETA and similar groups will protest and attempt to destroy your facilities if they hear of your experiment. You must take steps to prevent gossip.

OTTOMH I’m guessing that building a clean room and airlock will be at least $50,000. Windows and Mac software for reading EEG signals is available to the general public. This will cost between $500 and $5000 depending on the complexity of the software, the number of channels it can simultaneously monitor, the quality of the static (or artifact) filtering, etc.

DocCathode wrote

Could you give me some more detail?
What sort of nutrients?
What equipment is required to put nutrients and oxygen in and remove the wastes?
Can you give me brand names of equipment or links to places that sell this sort of thing?

This is an excellent point and I appreciate it.

Colibri or one of the other biologists can give you specifics. The basics are saline and glucose.

I’m not sure whether there is standard equipment that could be used for that. There are plenty of places to get equipment designed to deliver nutrients and oxygen to the bloodstream. A neural web in solution has no bloodstream. There are plenty of places to get equipment designed to oxygenate water. But, I doubt the gurgly thing in the aquarium is suitable for this job.

I have sent a link to this thread to a biochemist friend. I know he has lab experience with mammalian cell cultures. However, the next month will be busy for him and I don’t know when he can get back to me or how much time he’ll have to answer my questions.

Do you mean this: http://www.napa.ufl.edu/2004news/braindish.htm ?

A really quick search on PubMed didn’t turn up anything published by DeMarse on the subject in a peer-reviewed journal.

DeMarse has published about the tissue culture technique he uses though: http://tinyurl.com/aunhn .

Personally, as an advocate of animal experimentation, I would be horrified to learn of an unskilled person conducting this kind of experiment at home. In the UK animal experiments require licencing and approval before they can be conducted by a qualified person in an accreditied facility.

I’m no expert, but I’d have thought that the quickest, simplest, and cheapest method for using rat brain tissue to control a flight simulator would be to keep the rat in one piece and train it to use the joystick.

I can’t imagine going the brain-in-a-jar route without at least one hunchbacked assistant.

Brain Guy? Is that you?
Is Professor Bobo with you? How about Pearl?

You need a refill?

Szlater wrote

Thank you very much, Szlater. This text was extremely valuable, both in answering many questions and in uncovering my next level of research required to make this happen.

As a result of the work that Mssrs Potter and DeMarse discuss in this text, I don’t need to worry about some of the things that DocCathode was concerned about.

As a summary for those that might be interested, here’s what I’ve got so far:

I need access to a clean room to do the initial disection and setup of the petri dish.

The petri dish needs to have electrodes embedded in the bottom of it.

The rat brains to use are embryonic. I need a rat pregnant with 18-day old fetuses. It’s euthenized with CO2, embryo removed and cortex microdissected. Cortex pieces are digested in papain solution, and then deposited onto electrodes in petri dish.

The medium (which contains the food for the cells) is an off-the-shelf product called “modified Eagle’s medium” with some (10%) equine serum added. This is added to the petri dish. No additional things are added to the medium, such as antibiotics.

The lid of the petri dish is the “special sauce” of the project, and is really the point of the text that Potter/DeMarse were writing about. Traditionalally, the petri dish lid is lose-fitting, which had the benefit of allowing O2 and CO2 to equalize with the outside, which kept the medium oxygenated and kept the pH in check. Unfortunately, this also allowed water vapor to escape and air impurities to enter the dish, which meant live cultures couldn’t be kept for more than a month. Instead, I’ll use a fluorinated ethylene-propylene membrane, which allows O2 and CO2 to pass through it, but is relatively impermeable to water vapor. Potter/DeMarse claim they’ve kept cultures going for over a year with this technique.

The petri dish(es) are kept at a constant temperature (35 C), in an incubator. Because of the special petri dish lid, no humidifier is required.

Feeding is simply completely replacing the medium once per week in a laminar flow vent. Potter/DeMarse say that traditionally, one would replace half the medium in each feeding, but this was because evaporation caused the old medium to be denser in the non-water ingredients than the new, potentially resulting in shock upon complete replacement. But the hydrophobic petri dish lid alleviates this problem. They say they would’ve preferred a continual feeding system, but that hasn’t been necessary.

Any more thoughts or details by those that have been around this sort of thing are very much appreciated.

That’s what everybody always thinks, but then you have to consider the expense of building the little rat-sized simulator, and hiring an interpreter, and getting a little aviator’s outfit with the little tiny goggles, and then the rats, you know, try to TAKE OVER THE WORLD!

The petri dish is your best bet.

Brain, brain, what is brain!?!?

Training a rat to fly is limited. At best, you end up with a trained rat. The brain in a dish approach may lead to organic computers. The goal here isn’t ‘rat flies plane’. It is ‘brain in dish is given a plane as a total prosthetic and can control every aspect of the plane as simply and automatically as standard rat brains walk, see, hear, regulate heartbeat and breathing etc.’

Don’t even attempt to grow a brain.

Meh, what’s the worst that could possibly happen?

DocCathode wrote

Yes; this is exactly my direction. For those that are interested, google on “brain computer interface”. Wikipedia has a good starting description here: http://en.wikipedia.org/wiki/Brain-computer_interface. Also, some may be interested to google the upcoming product “braingate”. It’s been trialed in two humans already who were paraplegics (no mobility from the neck down), but with the use of this technology were able to control things by thinking about it, such as a TV remote control, reading email and opening and closing a prosthetic hand.

That should’ve read "quadriplegic. Here’s one article on it: http://networks.silicon.com/webwatch/0,39024667,39129194,00.htm

Bill H: The article you linked didn’t exactly inspire my faith. The pub date is April 1st, 2005. Upon further review it seems real. The sci-fi-y-ness of the first one, with the April 1st date threw me for a serious loop.
Here is what convinced me. Just in case anyone else noticed.

There are many articles in many places. I just picked the first google hit I saw.

By the way, it turns out I spent 30 bones unnecessarily on the Potters/DeMarse doc from the link provided. It’s available free many places, including here:
http://www.ece.gatech.edu/research/neuro/groups/potter/papers/PotterSealedDish.pdf