Hans Clevers: Extra footage 2

Hans Clevers gives a tour in his lab at the Hubrecht Institute, including images and explanation of organoids with PhD student Frans Schutgens; 3D images of a liver organoid; experiments with zebra fish.

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00:00:01 Part of the work is done in offices, with computers, but the vital part remains handwork.
00:00:09 We work in various laboratories...
00:00:12 This lab, I will open it...
00:00:16 Is entirely devoted to cultivating human tissue, the tiny organs.
00:00:23 The environment is completely sterile, hence the lab coats...
00:00:26 (interviewer) The organoids? Can we see them in this lab?
00:00:29 Yes, although I am not sure if we can see them right now... shall we just go in?
00:00:32 (interviewer) Yes please
00:00:42 Do you have something, an organoid, that we can see on the microscope?
00:00:48 (student) Yes
00:00:51 The cells are kept cool, so as to limit their growth, the cabinets are dust-free.
00:00:59 They are actually borrowed technique from space travel sciences.
00:01:05 The air is being filtered, because the bacteria in it fall on the cultures and kill them.
00:01:13 You can also see the gloves, lab coats...
00:01:22 This is a kind of modern Petri dish, actually 12 small Petri dishes on a plate.
00:01:30 They contain the nutrients, like carbohydrates and fats, and the grow factors we add.
00:01:43 If you do that correctly you see the organoids growing. We split them manually so that they continue growing.
00:01:56 This splitting is carried out every week, right?
00:01:59 (student) Yes.
00:02:01 These are kidney organoids?
00:02:04 Yes, tiny kidneys.
00:02:05 (interviewer) Is this a potential kidney?
00:02:08 So far it is just kidney cells, a kidney is an incredibly complex organ.
00:02:11 It has multiple layers of cells
00:02:17 We have just started this... have you figured out yet what type this is?
00:02:25 (student) This is mainly the functional tube part of the kidney.
00:02:30 The other part is still in development, we are still working on that.
00:02:37 This is nevertheless an important part of the kidney, and we can cultivate it quite well.
00:02:44 A kidney is very complex, we probably have to cultivate several components independently.
00:02:49 When we put everything together, my prediction would be...
00:02:52 That nature works in a way that instigates the spontaenous generation of a kidney.
00:02:56 We are not there yet, but the great thing is, we can cultivate these cells from urine.
00:03:00 There are always kidney cells in urine. You can use the urine from patients and healthy persons.
00:03:13 (interviewer) So this is alive?
00:03:16 This is growing, yes, but it is now cooling down slowly.
00:03:19 However, at a temperature of about 37 degrees celsius, you can witness the cell division.
00:03:27 Tomorrow this organoid is probably too big, so we then have to fragmentize it.
00:03:34 So it will fragmentize, but thereafter reorganise itself, become an organoid, and grow again.
00:03:38 There is no end to this process. If we know the initial conditions, which are different for every organ...
00:03:45 It is a bit trial-and-error to come to know the conditions, but you can repeat this endlessly.
00:03:53 You can do it 2-3 times a week. Some organoids grow in tenfold measure, so if you do that for a year...
00:03:59 You will have a universe full of intestines, or prostates, perhaps something you would not like...
00:04:06 We all learned that there is a limit to cell division. Normal cell division has a limit...
00:04:11 Because we think that endless growth equals cancer. This appears not to be the case.
00:04:18 How old is this culture?
00:04:20 (student) This one is 4 months old.
00:04:23 Growing continuously.
00:04:25 (interviewer) But how big is this culture in reality?
00:04:37 (student) This is about one milimeter, actually.
00:04:44 You can see them with the naked eye.
00:04:54 If you dare to lift the cover... bacteria and dust will fall in the dish, that is really bad.
00:05:01 You can see little globules, that is where these structures are in, there's a little condense so a bit hard to see...
00:05:15 (interviewer) You are holding a small box with life!
00:05:18 (student) That's right.
00:05:28 (student) So I have to look trough the microscope as I usually do?
00:05:57 Here are thin slices of organoid tissue, packed on small glass.
00:06:06 There is not that much to see, but there is a variety of ways, colour tagging methods...
00:06:14 To make a variety of things visible in the tissue.
00:06:17 This is classic stuff, what happens in a hospital, for example.
00:06:25 Purple is a general colour, different tints of purple reveal inflammations, normal tissue, viruses, and so on.
00:06:35 (interviewer) Can you identify these things as well?
00:06:37 For intestines - this is from a mouse, I can in this case, but pathologists know hundreds of diseases...
00:06:47 I have to look up things and consult others. Also, mice are not humans, so its different.
00:06:56 This is very important in a lab: music. There are endless discussions...
00:06:59 Music on? Music off? What kind of music? We now have two labs: one without and one with music.
00:07:07 This is also quite standard. With the help of an electromagnetic field and a fluid...
00:07:18 You can transfer tiny bits of DNA from the one side of the device to the other.
00:07:24 Bigger pieces tend to transfer slower, that is how we separate bits of DNA.
00:07:29 Think of typical images of forensic DNA research.
00:07:34 Producing a lot of DNA, cutting it into pieces, and comparing the pattern to the DNA of the potential culprit.
00:07:43 It is just basic molecular biology.
00:07:50 (interviewer) You do not have to show everything, only the things vital for your reserach.
00:08:02 This is the molecular biologist's essential tool. They are property like a pistol, or violin. It does the work.
00:08:10 P20. You set it, and basically what you do is extracting and injecting fluids.
00:08:17 That is the way to design every step in the DNA.
00:08:23 It happens always in this kind of tube.
00:08:30 If I want to cut the DNA, I add an enzyme, precisely on one location, which cuts it.
00:08:35 Then you have two pieces, which I can mix, add, you name it.
00:08:40 This is the core of molecular biology.
00:08:48 This is the microscope and we see the organoid sample on the screen. You see lasers, cabinets, and so on...
00:08:55 Biologists never build their own devices. Physicists do. We work with industry and usually buy devices.
00:09:03 This is beautiful. Can you turn it?
00:09:12 This is the size of a normal liver.
00:09:21 Every red area is one cell. You can see the nuclei.
00:09:29 Blue is a nucleus, purple is the boundary of the cells.
00:09:35 In all of these cells, there is at least one nucleus.
00:09:43 It is a bit difficult to see... can you see all the nuclei?
00:09:50 (student) It is difficult. Unfortunately, we don't really see the boundaries between the cells.
00:10:05 The flipside is beautiful, with the nuclei...
00:10:14 A liver has two cell types: we colour them red and green, and we see this liver generates them both.
00:10:21 (interviewer) But you add these colours.
00:10:25 Yes, but the possibilities are endless. It depends on what is informative.
00:10:40 The student tries to make much more tissue in three dimensoins. He uses a nylon threads...
00:10:47 The nylon meshwork apparently enhances the growth of the tissues quite well.
00:10:57 Together we thought of implementing the meshwork in patients, but made from biodegradables.
00:11:06 It may support this particular tissue structure. We ordered it, we hope that it works out for us.
00:11:15 What you see is a plug, it is only half a centimeter big, we could implement it in a liver.
00:11:21 Probably it will grow quite easily from there. We see that happening in mice already.
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