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Attila writes in with a how-to on isolating solate amniotic stem cells from a placenta, at home – this should generate a lot of conversations… check it out, the complete steps and discuss!

“The placenta is a very valuable human tissue, although the proper recycling of it is not placentophagy, but to isolate stem cells from its amnion layer, and store them for later regenerative purposes for the whole family. Human amniotic epithelial cells (HAECs) from the placenta are alternative replacements of human embryonic stem cells and have the potential to differentiate to all three germ layers in vitro. These cells are very close to those earlier and broadly multipotent amniotic fluid-derived stem cells, that made the big buzz lately on the web, published by De Coppi, Atala et al. in Nature Biotechnology. Here I would like to show, although I do not provide any warranty and can not give any guarantee, that isolating stem cells from the placenta is not more difficult than making a steak and with proper preparation, investment and timing you can do it even at home or in a rent lab. The process is ethically non-controversial since the placenta is usually discarded away after birth. Today, stem cell therapy is just a promising possibility, but in the not so distant future, self-aware citizens may manage their own stem cells, grow them in the garage, and store them in the fridge. If so, it could be a form of autonomous medical self-insurance. We are at the dawn of the biodiy movement backed by open source science, for the people. Make Backyard Biology issue was used, because it invented the basic language of bioDIY or home or garage biotech. Here is the algorithm at the cartoon and below are the detailed, although not self-including textual protocol. More details will come later, if asked.”

The rest of the image(s) and information after the jump….Link to full sized image – here.

1. Googlise human amniotic stem cells and the potential of stem cell therapy. It is not fact but a promising possibility.

2. Consult with some expert about the whole field (you can skip this step, if you are self-assured).

3. Talk to the pregnant member of the family (or acquaintances of yours) months before birth, and convince her to store the amniotic cells at home.

4. Consult with her gynaecologist months before birth and convince him to put the placenta under the birth into a sterile flask full with ice. You can put it into Phosphate buffered saline solution (PBS, preparation here) with some antibiotics (Penicillin-Streptomycin) for the sake of sterility.

5. Technical preparations (I did not calculate the exact amount of money, which is needed fro the adventure, but it’s around some thousands of dollars, and that could cheaper than collect the cells via a commercial way): Set up a sterile hood at your garage. You can make one out of a household air purifier, for that see Home Mycology Lab by Philip Ross, in Make Backyard Biology issue, page 102. Rent or buy a normal light microscope (10x resolution will be enough), a centrifuge (1000rpm), and buy a liquid nitrogen refrigerator.

6. When the placenta is in your hand, process it within 4 hours. Use sterile gloves.

7. Put the flask with the placenta under the sterile hood. Take a sterile scissor and carefully cut the outside epithelial layer off. The more you cut, the more stem cells you get. The amnion layer is mechanically peeled off the chorion.

8. Wash the amnion in Phosphate buffered saline solution (PBS, preparation here) in several times (8-10X) to remove blood.

9. Mince the tissue thoroughly with sterile scissor.

10. To release amniotic epithelial cells, incubate the minced amnion membrane with trypsin (0.05%) for 10 minutes at 37°C. Take out the digested tissue from trypsin after 10 minutes, and discard the cells from this digestion to exclude debris. There are different kinds of trypsinization protocol, I follow here Miki et al.

11. Treat the remaining tissue in another tube of trypsin (0,05%) for 20 minutes at 37 °C (do this step once more if necessary to collect more cells).

12. Pool the cells from second (and third 20 minutes) digests.

13. Pass cell suspension through a 100 μm cell strainer.

14. Fuge the filtered cell suspension for 8 minutes at 1200 RPM (150-200g), room temperature.

15. Wash the cell pellet with PBS and fuge again.

16. Count the cells with a hemocytometer and it is advisable to determine the viability of the cells by exclusion of trypan blue dye, which is based on the principle that live cells possess intact cell membranes that exclude certain dyes, such as trypan blue, whereas dead cells do not. From one epithelium you can get as many as 10-60 million stem cells.

17. Prepare freezing medium. The freezing protocol is from the best lab manual, At the Bench: A Laboratory Navigator by Kathy Barker: 1ml/aliquot plus 10%. Freezing medium typically contain regular culture media, 10-20% serum, and 5-10% glycerol or DMSO.

18. Each ampoule will take 1×10(7) cells (or between 4×10(6) and 2×10(7) cells) in 1ml of medium.

19. Resuspend the pellet in freezing medium by pipetting gently.

20. In order to freeze the cells gradually and safe place the ampoules in -60°C or less and leave them there for 16-24 hours.

21. Put the aliquots with the cells in liquid nitrogen and store them. As Charles Platt says in Life and Death at Low Temperature (page 56): “liquid nitrogen is available in most urban areas (search for “liquid gases”), and it is generally inexpensive. It is nontoxic, but must be handled with caution, since its temperature of -196°C can cause serious injury to any exposed human tissue. Always wear heavy gloves and eye protection.”

22. Give the freezed cells to doctors, when it is needed for repair after X years.

good: frozen

Phillip Torrone

Editor at large – Make magazine. Creative director – Adafruit Industries, contributing editor – Popular Science. Previously: Founded – Hack-a-Day, how-to editor – Engadget, Director of product development – Fallon Worldwide, Technology Director – Braincraft.


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Comments

  1. herr_prof says:

    ewww!! RAD!!!!

  2. cbleslie says:

    Can I still make the sandwich?

  3. Archvillain says:

    Very cool.

    It looks like there is plenty left over, so I’m guessing you can still make the sandwich cbleslie :-)

    “The process is ethically non-controversial since the placenta is usually discarded away after birth.”

    That might sound like common sense, but excess embryos from fertility treatment are likewise usually discarded away after birth, yet by directive coming down direct on high from the Prez Himself, you Can’t Touch That.

  4. Daenris says:

    That might sound like common sense, but excess embryos from fertility treatment are likewise usually discarded away after birth, yet by directive coming down direct on high from the Prez Himself, you Can’t Touch That.

    Indeed, but the placenta isn’t an embryo :)
    And hopefully no one is arguing that the placenta is a life and whatnot.

  5. Sally599 says:

    Um hello..how many people have a whole lab set up in their home? Aside from the $$$to get a centrifuge etc, you’d need $$$ for liquid N2 which is still going to cost you a couple of hundred every few months to fill the tank. Plus I would never trust a sterile hood made from an air purifier, or just adding antibiotics rather than 0.22 uM filtering your PBS or even autoclaving. Ick Ick Ick, leave it to the pros. The feds won’t fund stem cell research but several states are and so are private companies. Besides there are lots of other countries that would love to be pioneers in stem cell research, if you’ve got the money to blow to set up a whole lab in your house why not set up a small company in a third world country?

  6. dculberson says:

    Note to self: don’t read Makezine Blog while eating pizza.

  7. trebuchet03 says:

    “yet by directive coming down direct on high from the Prez Himself, you Can’t Touch That.”

    That just means any medical breakthrough using that route will happen in a nation other than the United States :P

    “Um hello..how many people have a whole lab set up in their home?”

    20+ years ago — that sentiment would be — “How many people can afford a whole computing set up in their home? (and have space)” If there’s a demand — there will be a whole lot of companies explaining why their equipment is not only cost effective, but better than their competitors :P

  8. fstedie says:

    I agree, this is much more than hacking a speak’n spell. More like a biology grad student….

  9. Archvillain says:

    how many people have a whole lab set up in their home?

    Thanks to the private sector biotech boom, company liquition, the magic of ebay, and onsellers, you too can enjoy the spoils of commercial laboritory equipment at pennies-on-the-dollar prices :-)

    And if you cut out the middle man, the stuff is even cheaper :)

  10. BrK says:

    I agree, this is much more than hacking a speak’n spell. More like a biology grad student….

    Sure, but haven’t we seen enough of circuit bending and felt-covered whatever already?

    I may never do this sort of thing, but at least it’s something new and more Make-y than a minor variation on an already over-publicized thing (bending).

  11. philliptorrone says:

    BrK – compared to all the gadgets and other over-publicized things you see on 99% of the tech sites i don’t think felt and circuit bending coverage is overly promoted here, but that said – it’s what our makers are doing and sending us.

  12. Sally599 says:

    Its true that you can buy cheap equipment on E-bay, but there is always a catch. A young assistant professor down the hall bought an inverted fluorescent microscope on E-bay, and while it works the light bulb burns out every 24 hours instead of 6 months. These are propietary mercury bulbs and cost around $200 a piece so she’s paying full price for the scope in the end.

    As far as labs being common in the home, I think its the opposite trend from computers. Labs were very common in the homes of early scientists, Pastuer etc, but its gone the other way as the need for specialized and diverse equipment has increased. Often we need to go to other labs or core facilities to use equipment because we can’t afford everything.

    Still, I think its a fun post but really who wants to store a biological and chemical(N2) hazard in their home. The feds would be on your doorstep investigating the possibility of bioterrorism before your equipment could ship from Fisher.

  13. hammerthumb says:

    Sally599: “Still, I think its a fun post but really who wants to store a biological and chemical(N2) hazard in their home.”

    Someone who is a sole proprietor of a private research company that has converted their garage into a laboratory? Someone with lots of money and curiosity and no connections to academia?

    As an aside, I store yeast and CO2 for my home brewed beer without a problem.

    “The feds would be on your doorstep investigating the possibility of bioterrorism before your equipment could ship from Fisher.”

    It’s not illegal to isolate and store placental stem cells. Calculate legal fees into the cost of your lab and then get a consultation with a suitably experienced lawyer. Provided you’re not an actual terrorist, I’m sure there are things you can do to keep the Feds informed and minimize the disturbance to your laboratory.

  14. Sally599 says:

    Yeast and CO2 are not serious biological or chemical hazards. Human tissue of any type is a biological hazard because of the possibility of carrying HIV or any number of viruses the person you get the placenta from may not even be aware of. You would have to test for all of these things before ever using the cells. Personally I wouldn’t want to mash up and therefore possibly aerosolize potentially infectious material. Usually tissue for lab research is give a test for HIV/Hepatitis etc before people in a lab ever handle it. The hood would protect you to some degree but when working with an unknown you’d really want to use at least level 3 safety precautions.

    The N2 is a chemical hazard not so much from the whole heavier than air problem which could suffocate you which would also be possible with CO2 but from the severe burn problem and from the explosion problem. From a burn perspective…if you had the right protective equipment, gloves etc it probably wouldn’t be a problem for an adult—when I picture an actual house I picture kids running around getting in trouble. Liquid N2 is really fun to play with. Now the explosion problem. Those little cryotubes can explode. They are small plastic tubes and are not reinforced. They are meant to be used only in the gas phase of N2 and if you accidentally immerse them in the liquid N2 they pose an explosion risk as they are warmed to room temp. We usually wear face shields, lab coats, heavy gloves, etc—aside from the flying plastic shards you are also getting biological sample sprayed everywhere…if you haven’t tested for Hepatitis, etc. in the sample then you’ll be making a quick trip to the doctor because you may have exposed yourself.

    I’m not saying it can’t be done…but you had better know what you’re doing and there are easier ways than this.

    As far as the feds, whats legal and what keeps you under the radar are two different things. Personally I don’t want to be put on the list for “special” security checks at the airport.

  15. Pekar says:

    Wort zu deiner Mutter!

  16. Trypan blue poses a health hazard (it can be irritant to the skin and is a potential carcinogenic). I’d say a more adequate choice is Erythrosin B, which can discriminate live vs dead cells and poses no health issues (it’s even used as a color additive in foods). Check out the comparison in the hemocytometer blog.

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