information about stem cells

shall we have a look at your baby? - you're 27 weeks now, aren't you?- yeah.- okay. oh, that's a nice one, isn't it?that's always good, isn't it? birth defects represent one of the major causes of morbidity and mortality in children. there's about three in every 100 babies

which are born with a major birth defect. we are faced many times with a situation in which we don't know what to do for these babies. - it's a girl.- it's a girl. i collaborate with paolo de coppi.i look after the mums while they're pregnant. paolo visits the fetal medicine unit,and he sees the mums if the baby has an abnormality because he looks after the babywhen they're born. and we're trying to develop

treatments using stem cellsin the amniotic fluid around the baby. stem cells have represented a major breakthrough because it's the possibility of growing cellsoutside the body to make a repair to the children once the cells have been built in the laboratory. so what we have developed in the last few years and this has been initially the work we have done

with antony atala at the wake forest institute of regenerative medicine and is the possibility of deriving stem cell from the amniotic fluid. these are not embryonic stem cells but are neither adult stem cells. they have characteristics that are in between the two cell types, and for that, they present a bigadvantage for therapy. these cells would represent an ideal source

for building organ and tissue that are missing in the foetus. and this is because we can predict and diagnose very accurately these diseases before the foetus is born. once this diagnosis is made, however, we have about 20 weeks of gestation in which we can plan the engineered organ to be built outside of the baby,

that then can be eventually implanted once the baby is born. so we can correct that defect at birth using his own cells. here is where we do receive the samples from anna's. once the cells are isolated from the amniotic fluid,they can be easily expanded in this incubator

and eventually engineered in a three-dimensional structurethat can mimic the organs that the baby is missing. these organs can be expanded and grown in these bioreactors. we haven't got any treatments that will actually work yet. but we're working on this whole area. for instance, the mum that we scanned earlier onhas got a completely healthy, normal baby as far as i can tell on the scan. we're not looking to treat anybody right now.but we are trying to develop new

treatments which will improve already existing treatmentsor to develop completely new therapies, using the stem cells that are in the amniotic fluid. another situation in which we can intervene even before birth - we can use the same cells to treat the foetus. so if we know that the foetus has some malformation that can be corrected before birth,

we can use his own cells to, for treatment. so we can culture his or her cells outside the womb and expand them and eventually correct, for example, the gene that is altered or missing, using gene therapy technique. and we can inject back the cells

into the foetus before birth. and so somehow improving his option of life

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induced pluripotent stem cells

this video will show you how to coat cultureplates with vitronectin, so you will be ready to transfer newly forming ipsc colonies orfrozen cultures to essential 8â„¢ medium. before using the essential 8â„¢ complete medium,you'll need to prepare vitronectin-coated plates. the optimal working concentrationof vitronectin is cell-line dependent and must be determined empirically. we recommendedusing a final coating concentration of 0.1–1.0 âµg/cm2 on the culture surface, dependingon your cell line. we routinely use vitronectin at 0.5 âµg/cm2 for human psc culture. prior to coating culture vessels, calculatethe working concentration of vitronectin using the following formula and dilute the stockappropriately. the product manual for vitronectin

contains a list of culture dish surface areasand volume of vitronectin required. as an example,, to coat a 6-well plate at a coatingconcentration of 0.5 âµg/cm2, you’ll need to prepare 6 ml of diluted vitronectin solution(10 cm2/well surface area and 1 ml of diluted vitronectin/well). upon receipt, thaw the vial of vitronectinat room temperature and prepare 60âµl aliquots of vitronectin in polypropylene tubes. freezethe aliquots at –80â°c or use immediately. to coat the wells of a 6-well plate, removea 60âµl aliquot of vitronectin from –80â°c storage and thaw at room temperature. you’llneed one 60âµl aliquot per 6-well plate. add 60 âµl of thawed vitronectin into a 50-mlconical tube containing 6 ml of sterile dpbs

without calcium and magnesium at room temperature.gently re-suspend by pipetting the vitronectin dilution up and down. this results in a working concentration of5 âµg/ml, a 1:100 dilution add 1 ml of the diluted vitronectin solutionto each well of a 6-well plate. when used to coat a 6-well plate (10 cm2/well) at 1ml/well, the final concentration will be 0.5 âµg/cm2. if you’re using a different type of culturevessel, refer to the vitronectin product insert for detailed information on volumes. incubate the coated plates at room temperaturefor 1 hour.

note: the culture vessel can now be used orstored at 2–8â°c wrapped in laboratory film for up to one week. do not allow the vesselto dry. prior to use, pre-warm the culture vessel to room temperature for at least 1hour.

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human stem cells

an international team of researchers has developedan artificial version of the human midbrain using stem cells. the team's creation will allow for more extensiveresearch and drug testing,... and could have broad treatment implications -- especiallyfor degenerative disorders involving the motor system. park jong-hong explains. the breakthrough could eventually be life-alteringnews for patients of parkinson's disease. the leading degenerative disorder of the centralnervous system is a condition stemming from the midbrain, which is in charge of motorfunctions that control auditory and eye movements,

vision and body movements. the midbrain contains special neurons thatproduce dopamine, and the disease develops when the number of neurons decreases. with the breakthrough, scientists have createda miniature version of the midbrain, which they hope will shed light on exactly how parkinson'sevolves and lead to a cure for it and other aging-related brain diseases. while miniature versions of the brain havebeen developed before, this one is the first of its kind. it is a three-dimensional miniature with tissuesthat were grown in a laboratory using stem

cells cultivated from human blood, and itcan be used in a variety of drug tests instead of in experiments on actual patients. the medical community is abuzz about the possibilitiesfor research and treatment the breakthrough will have. the joint study was conducted by an internationalteam led by professor shawn je from duke-nus medical school and a*star's genome instituteof singapore. their findings were published this month inthe journal cell stem cell. park jong-hong arirang news.

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human mesenchymal stem cells

- [voiceover] so, let megive you an analogy, here. when you were still anadorable little baby, you were just bursting with potential. you could decide to be a pilot, or a doctor, or a journalist. you had the potential to specialize into all sorts of different careers, and as you got a bit older,you got more and more committed down a certain pathway,

and the decisions that you made moved you further and furtheralong this pathway, right? well, it turns out that stemcells operate in a similar way, going from unspecializedto more specialized as they get older. so, let me show you what i mean by that over the course of this video. and let's actually startback at the zygote, here, the cell that resultswhen sperm and egg fuse

because that's really where our stem cell story kinda begins. so, the zygote starts to divide, right, by mitosis until it reachesthe blastocyst stage, this hollow ball of cellshere is called a blastocyst. and here, things start to geta little bit more interesting. so, in a blastocyst, there'sthis little grouping of cells down in here, referred toas the inner cell mass. and this is a really speciallittle bunch of cells

that go on to become the embryo. so, these are called stem cells. and what they can do as stem cells is they can specialize intoseveral other cell types. so, we actually call thempluripotent stem cells. pluri meaning several and potent referring to these stem cells' ability to actually do this differentiation. so, during development,these inner cell mass

pluripotent stem cells can differentiate into any of the more than200 different cell types in the adult human body whengiven the proper stimulation. so, it's kind of incredible to think that every single cell in your body can trace its ancestry back to this little group of stem cells, here. and actually, if you ever hear anyone talking about embryonic stem cells,

these are the ones they're referring to, these icm stem cells. so, is this the only placewe can find stem cells, here in the developmental structures? we used to think so, but, it turns out that in mammals, there aretwo main types of stem cells. embryonic stem cells that we just saw and somatic stem cells whichare found in every person. so, the embryonic stem cellsare used to build our bodies,

to go from one cell totrillions of specialized cells, and the somatic stem cells are used as sort of a repair system for the body, replenishing tissuesthat need to be replaced. and they can't repair everything, but, there's a lot of every day repairs that can happen because of our stem cells. so, in skin, for example... this outside layer is the partof our skin that we can see

and that we can touch, right? and it's made of these waterproof, pretty rugged epithelialor skin cells and interestingly, althoughthey are pretty rugged, you're constantlyshedding these skin cells. they actually just sort offall off or get rubbed off during every day activities like when you're putting your clothes on. and then, the ones from underneath them

just sort of move up and take their place. so, you shed them and you lose almost 40,000 of them per hour. so, if we wanna have anyhope of keeping our skin, we kinda need a way toreplace these cells, and that's where stem cells that live in our skin come in. actually, our skin cells are shed and replaced so often,that it only takes a month

for us to have a completely new skin. like, literally onemonth, entirely new skin. it's outrageous. anyway, deep within our skin, there's this layer of stem cells called epidermal stem cells, and their job is to becontinually dividing. so, you can see themdividing, here, dividing, dividing, dividing, and makingnew skin cells that go on

to migrate upward as themultiple layers of our skin. and their goal is to eventually replace these ones up here on the outside that get damaged or worn out and fall off. so, it's this kind of activity here which show off our stem cells' role as our regenerative cells. now, lemme just highlighta few differences between our mature skin cells over here

and our stem cells down here. they are very different. mature cells are notthe same as stem cells, and this principle goesfor really any mature cell versus any stem cell. so, the mature cell isalready specialized, it already has a really specific function. for example, our outer layerof epithelial cells, here, they have a protective function

against the outside environment. and, you know, just thinkingof other adult cell types, right, like muscle cellshave a contractile function, and neurons have amessage sending function, and bones have a rigidstructural function. so, all these adult cells are already nice and specialized, they'vegrown up and decided what they wanna do for a living, whereas, stem cells arenot like that at all.

stem cells are unspecialized. but, they still have areally important job, which is to give rise to ourmore specialized cell types, like these cells here, okay? and, actually, in order tobe considered a stem cell, and this goes for theembryonic stem cells we met previously and the somaticstem cells we're meeting now, to be a stem cell, you'd need to possess two main properties.

the ability to self renew,meaning you can divide and divide, and divide, but, at least one of your resultingcells remains a stem cell, it remains undifferentiated, and you'd need to have a high capacity to differentiate intomore specialized cells when the time comes. so, remember, this is also referred to as having some degree of potency.

and there's actually a few different types of stem cells, and someof them can turn into more types of cells than others. some are more potent than others. so, this epithelial stem cell we saw here is actually one of the lesspotent types of stem cell. in other words, thesestem cells can only divide and specialize into more epithelial cells. so, they're our source ofepithelial cells, sure,

but, only epithelial cellsand not any other cell type. so, we call them unipotent,referring to their ability to only create one type of cell. but, lemme show you another example here of a multipotent stem cell. let's look at this guy'sfemur, his thigh bone, which is where our blood cells are made inside bone marrow in our bones. so, you might know thatour red blood cells

have a life span of about four months. so, that means that we needto be constantly replacing our red blood cells orwe'll run out, right? well, in our bone marrow,we have what are called hematopoietic stem cells, which are our blood making stem cells. and these are pretty special, they're multipotent stem cells, which means they can giverise to many types of cells,

but, only ones within a specific family. in this case, blood cells,and not, for example, cells of the nervous systemor the skeletal system. so, our hematopoietic stem cells are always busy churningout new blood cells, red blood cells to carry oxygen for us, and white blood cells to keep our immune system nice and strong. and for a more clinical example,

with blood diseases like leukemia, certain blood cellswill grow uncontrollably within a patient's bone marrow, and it actually crowds out their healthy stem cells, here, from being able to produce enough blood cells. so, as part of treatment,once the leukemia cells are cleared from the bone marrowwith, usually, chemotherapy or radiation, doctors can actually put

more hematopoietic stem cellsback into the bone marrow that then go on to produce normal amounts of blood for the person again. so, this is probably the most common use of stem cells in medicine as of now. and you can actually findthese multipotent stem cells in most tissues and organs. so, for example, we havemultipotent neural stem cells that slowly give rise to neurons

and their supporting cells when necessary. and we have multipotentmesenchymal stem cells in a few different places in the body that give rise to bonecells and cartilage cells, and adipose cells. so, you might be wonderingafter seeing our epithelial and our hematopoietic stem cells dividing, why aren't these cells beingused up as they divide? and that's a really good question.

so, stem cells havetwo mechanisms in place to make sure that theirnumbers are maintained. so, their first trick isthat when they divide, they undergo what's calledobligate asymmetric replication where the stem cell dividesinto one so called mother cell identical to the original stem cell, and one daughter cellthat's differentiated. so, then, the daughtercell can go on to become more specialized while the mother cell

replaces the stem cellthat divided, initially. the other mechanism is calledstochastic differentiation. so, if one stem cellhappens to differentiate into two daughter cells insteadof a mother and a daughter, another stem cell will notice this and makes up for the lossof the original stem cell by undergoing mitosis andproducing two stem cells identical to the original. so, these two mechanisms make sure

their numbers remain nice and strong. so, we've looked at embryonic stem cells and we've looked at somatic stem cells. there's actually one more type called induced pluripotent stemcells, or ips cells. it turns out that youcan actually introduce a few specific genes intoalready specialized somatic cells like muscle cells, andthey'll sort of forget what type of cell they are,and they'll revert back,

they'll be reprogrammedinto a pluripotent stem cell just like an embryonic stem cell. and this is a huge discovery. i mean, the technique isstill being perfected, but, there's a lot ofmedicinal implications, here. for example, ips cellsare basically the core of regenerative medicine,which is a pretty new field of medicine where the goalis to repair damaged tissues in a given person by using stem cells

from their own body. so, with ips cells, each patient can have their own pluripotent stem cell line to theoretically replaceany damaged organs with new ones made out of their own cells. so, not only would apatient get the new organ they might need, but, there also won't be any immune rejection complications since the cells are their own.

so, there's still a ways to go here before this type of medicineis sort of mainstream, but, already, ips cells have helped to create the precursorsto a few different human organs in labs, suchas the heart and the liver. now, before we finish up here, i just wanna answer two questions that might have come up for you. so, one, what triggers ourstem cells to differentiate?

well, it turns out thatin normal situations, right, when the stemcell's just hangin' out, not doin' too much, it actually expresses a few different genes that helps to keep it undifferentiated. so, there are a few proteinsfloating around in the cell that prevents other genesfrom being activated and triggering differentiation. but, when put in certain environments,

this regulation can be overridden, and then, they can go on and differentiate into a more specialized cell. the type of which depends on what specific little chemical signals are hanging around in the stem cell's environment. so, for example, in the bone marrow, there are certain proteinsthat hang around stem cells and induce some to differentiate

into the specific blood cell types. and finally, what's all thisstuff you might have heard, maybe in the news, about cord blood? well, from cord blood,which is blood taken from the placenta and the umbilical cord after the birth of ababy, you can get lots of multipotent stem cells, and sometimes, some other stem cells that have been shown to be pluripotent.

so, this cord blood usedto just be discarded after a baby's birth, but now, there's a lot of interest in keeping it because now we know itcontains all these stem cells.

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how to get stem cells

i want to tell you one story. and the storycame from the early 1950s. a woman named wanda ruth lunsford, she was a scientist in newyork city and she published one paper, which turned out to be her only paper in scienceand she was actually pushed out of science. what she did was she took an old rat and ayoung rat, she put them to sleep and she tied their skin together. so after about a dayor so their blood supplies joined. well, several weeks later she looked and in that old ratthere were new neurons growing in the brain, the heart beat stronger and the muscles werebigger. the gray hair turned brown again. she claimed she reversed aging. people callher dracula, frankenstein, all kinds of crazy names.

well, earlier this year three separate laboratoriesat harvard, stanford, university of california san francisco repeated the experiment andit worked. and what they showed is at age 25, in you and i, our stem cells go to sleepand get turned off. and proteins, from young mice in this case or young humans, can turnthem back on again. and when these stem cells get turned back on new neurons can be grown,repair happens much quicker in tissue. we all see that. our child breaks his leg heor she is back walking again in a couple weeks. you don't even know what happened. your grandmotherbreaks her leg and it hits her quality of life the rest of her life. so there are clinicaltrials now using proteins that were found in young individual to try to stimulate bonerepair in the elderly who have fractures.

and so just like a diabetic requires a shotof insulin so that they can manage their sugar, going forward if you break your leg in theelderly we may just give you a shot of these proteins to turn back on your stem cells soyou can repair quicker. we're trying it in cancer because cancer inkids is about 90 percent curable. once you turn 25 that same cancer turns incurable.so maybe if i can convince the body it's younger i can have, or we as a science community canhave, a bigger impact on cancer. so i leave you with that bit of hope that aging is somethingthat may be able to be reversed, and not so that we can live till 150 but so that we canall live until our ninth or tenth decade without there being a decrease in quality in thoselast decades, because that would be the goal,

quality years till the end.

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how do you get stem cells

embryonic stem cells are a huge next stepfor medicine, but they’re mired in controversy. so what if you could have all the benefitsof regenerative medicine, without the thorny moral dilemma? hello science lovers, julian here for dnews.there’s been a huge breakthrough in medicine this week; induced pluripotent stem cellswere used in a tissue transplant for the first time ever. we did it! yeah!! oh am i goingto need to explain that? ok. first, let’s get everybody up to speed onstem cells. this is more important to you than you realize because you were at somepoint nothing more than 50 to 150 of them. you probably don’t remember, it was only4 days after you were conceived.

what makes embryonic stem cells amazing isthey are pluripotent, meaning they can become almost any type of cell. this means they canbe used to replace damaged tissue and may one day be an alternative to organ transplants.this is huge. in the united states an average of 79 people receive an organ transplant everyday, but another 18 people die waiting for a transplant that will never come. the 79 who do get a donor will have to takemedication to prevent transplant rejection for the rest of their lives, but even thenthe risk remains. that problem doesn’t exist with embryonic stem cells. so we can savethousands of people a year and improve the quality of life and lifespan of tens of thousandsmore. what’s the problem here?

well the only way to get pluripotent cellsis from embryos. which are pre-people. or current people, depending on where you standon the issue. obviously, there’s a heated controversy there and i’m sure the commentswill reflect that (be nice, guys). umbilical cord blood has multipotent stem cells, butthey’re more limited. and we all have adult stem cells but those are even less versatile.so there are no alternatives. until now. shinya yamanaka of kyoto university discovereda method to create pluripotent stem cells from adult cells. it’s like diet stem cells:all of the flavor with none of the calories. though “flavor” in this case means life-changingbenefits and “calories” means severe moral dilemma of using undeveloped humans in medicine.and for his breakthrough, yamanaka shared

the nobel prize for physiology in 2012. and now, here we are, september 2014, whereon friday, at 2:20 local time, japanese doctors stopped the macular degeneration of a 70 year-oldwoman by transplanting a sheet of retinal pigment epithelium cells derived from inducedpluripotent stem cells made from reprogrammed cells of the patient’s own skin! we didit! probably! now we wait and see if the procedure checks the macular degeneration without becomingcancerous or rejected. by the way the sheet of tissue used was just3 by 1.3mm. that’s about that big. and what a huge step that is. if you’d like to learn more about how youreye works and why the macula is important

to vision, check out this video. as a bonus,you can stare deep into trace’s dreamy, dreamy eyes. can you think of any creative potential usesfor induced pluripotent stem cells *cough* wolverine *cough*? let us know in the comments.hopefully it stems productive discussion. remember to subscribe for the latest in science,and i’ll see you next time on dnews.

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how do we get stem cells

this video will show you how to passage pscswith edta in essential 8™ medium on vitronectin coated plates. the following protocol uses a 6-well plate.if you’re using another type of cell culture vessel, please refer to table 2 on the writtenprotocol for volumes there are three major differences that you’llobserve with cells cultured in essential 8™ medium on vitronectin compared to other feeder-freesystems such as mtesr and stempro hesc sfm: • cells are typically passaged ~24 hourssooner than they would be with other feeder-free media.• passaging should take place when cells are at ~85% confluency. if cells are passagedwhen they are more than 85% confluent, the

health of the cells and final cell yield maybe compromised. • cells must be passaged in edta. collagenaseand dispase are not recommended. cells will reach optimal confluency typicallyevery four to five days. you should also split cultures or passagecells when psc colonies become too dense or too large or show increased differentiation; the split ratio can vary, although it’sgenerally between 1:2 and 1:4 for early passages and between 1:3 and 1:12 for established cultures.occasionally, cells will grow at a different rate and the split ratio will need to be adjusted. a good rule is to observe the last split ratioand adjust the ratio according to the appearance

of the psc colonies. if the cells look healthy,and the colonies have enough space, split your cultures using the same ratio. if thecolonies are overly dense and crowding, increase the ratio; if they are sparse, decrease theratio. newly derived psc lines may contain a fairamount of differentiation through passage 4. it’s not necessary to remove differentiatedmaterial prior to passaging. by propagating/splitting the cells, the overall health of the cultureshould improve throughout the early passages. prepare 0.5 mm edta by combining 50 âµl ofultrapureâ„¢ 0.5 m edta, ph 8.0 with 50 ml of dpbs without calcium and magnesium. filter,sterilize the solution. and store at room temperature.

pre-warm complete essential 8â„¢ medium andvtn-n-coated culture vessels to room temperature. aspirate the spent medium from the vesselcontaining pscs and rinse the vessel twice with dpbs without calcium and magnesium. add 0.5 mm edta in dpbs to the vessel containingpscs. swirl the vessel to coat the entire cell surface. incubate the vessel at room temperature for5 to 8 minutes or at 37 degrees celsius for 4 to 5 minutes. when the cells start to separateand round up, and the colonies appear to have holes in them when viewed under a microscope,they are ready to be removed from the vessel. note: in larger vessels or with certain celllines, this may take longer than 5 minutes.

aspirate the edta solution, and add pre-warmedessential 8â„¢ medium to the vessel. remove the cells from the well(s) by gentlysquirting medium and pipetting the colonies up. avoid creating bubbles. try to work with no more than 1 to 3 wellsat a time, and work quickly to remove cells after adding essential 8â„¢ medium to thewell(s), which quickly neutralizes the initial effect of the edta. some lines re-adhere veryrapidly after medium addition, and must be removed 1 well at a time. others are slowerto re-attach, and may be removed 3 wells at a time. collect cells in a 15ml conical tube. theremay be obvious patches of cells that were

not dislodged and left behind. don’t scrapethe cells from the dish in an attempt to recover them. add an appropriate volume of pre-warmed essential8™ medium to each well of a vitronectin coated 6-well plate so that each well contains2 ml of medium after the cell suspension has been added. move the vessel in several quick figure eightmotions to disperse the cells across the surface of the vessels. place the vessel gently intothe 37 degrees celsius, 5% co2 incubator and incubate the cells overnight. feed the psc cells with essential 8™ mediumbeginning the second day after splitting and

replace the spent medium daily. it is normal to see cell debris and smallcolonies after passage.

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