Introduction of Cell Culture Technology
Good. Evening and welcome. You, all to. This new course which, we'll be starting today on cell, culture technology. So. The. Name. Cell. Culture whenever it comes, across. People, have the first feeling that well it's a. Lab. Training, or a, kind. Of a. Technique. Which is used by most, of the labs. Who. Work at the interphase area biology chemistry and vanduul, sciences and. All. Other allied fields my, Medical Sciences say, for example, so. It's. Just a simple technique what most of the lipid kind of understand, that you know. Cells. Which are cultured, in and synthetic. Or artificial environment. At. Least that's what the commoner, or. You. Are still in graduate students, or undergraduate. Student, that's what they will answer. Why. We, really want to offer a course in this area. So. And. As a matter of fact when I floated this course. The. Refusal has me that that's a practical training, how, really, you want to. You. Know put across a course, which is mostly, a practical, training people have cell culture labs. So. For. Me. They. Put it across is. Though. The name is just, cell culture. But. There is a philosophy behind this whole thing. There. Are some. Basic. Rules and. Moreover. There. Is a, tremendous. Amount of, science and, art. Involved. In it and. If. Somebody. Who. Wants to do cell culture or. Wants. To learn cell culture. Appreciates. These. Basic. Philosophy. Or the paradigm. Or. The. Milestones, then. It will be easy for that individual, to design, the problems, better and. Most. Importantly. To. Appreciate. How, far they can go, say, for example in, mathematics, there are several techniques right or. In. Physics. There are several techniques. Similarly. In chemistry there are several techniques, say. Spectroscopic technique. But. We have course, the. Basic logic. Behind, the spectroscopy where there's the Raman spectroscopy, whether, it's a if, you, IR whether it is NMR. If. You know those at least the basics then, you know with, your problem, what. All you can derive using spectroscopy, or. Say, for example crystallography. Similarly. Several mathematical, tools, similarly. Like statistics, there, are these are tools but. If, you know the tools and the origin of the tools and the power of the tools it. Will help you to, become much more wiser in, designing. Your problem. Similarly. Cell, culture is. A. Tool but. Tool. Whose. The, journey. Will look back. Is now, more than hundred years old and. There. Are handful of books it, will come across and some of them I have already. Mentioned. Like yum freshness. Tissue. Culture book or. A. Few. Other books like Gosselin. Bankers. Neural. Cell culture book and, there. Are manuals, published, by. Companies. Like gyp. Go in with Trojan or, currently. Which is known as inveterate, life, science technologies. And. They. Are really nice guide, but. One thing which I, personally. Have. Felt is that. Somewhere. These. Books are more or less like a protocol, book, but. Cell culture as such is not just. A mere protocol, is definitely, there is a significant. Part of it which is critical, driven just. Like in molecular biology. You. Have many. Artists is manual. Meaning and goes through it okay but. It's still molecular, biology, as a. Subject. Has. Tremendous. Amount of chemical. Logics and unless, you understand that you will be working like a blind person, that's. Precisely, what happens, in cell culture, that. My. Young student, are. Young. Enthusiastic. Undergraduate. Or postgraduate. Just. Embraces, that this is technically, just let's follow, it without. Putting logic, in place that why, am i doing it. What. Is the basic reason behind it, so. In this course what, we try to do of course we will talk about the. Techniques but mostly. We will be talking about the philosophy and what. Drives us and what are the possible, options. Some. Explored, some, unexplored, as I. Mentioned, just a couple of minutes back. The. History of cell culture is. Now. More than hundred years old and. If. You look back. Like. If you are a good historian, or if. You are interested in scientific history. If. You look back the. Very first paper, which. At. Least known to us. Which. Were published in. This. Field. Where. Around. 1908. 1912. It. Is that back even much earlier, than the First World War what world wars started in, 1914. Right. So. This. Was the paper from Harrison, one of the pioneers and. What. He did for the first time he. Grew. Explant. Expands, means part. Of. Some. Animal. Systems body, on. A matrix, and the matrix was, a very interesting thing it. Was a spider net. He. Didn't know where to grow. So. If they break up this problem what.
He Was trying to do but. Before what, is trying to do. Let's. Go. Back a little so. If this field which, has its. First, set of publication, coming almost. 117. Years back. Then. For. Most what. We'll try to do we'll try to. Appreciate. This whole history of 100 years what. Has happened how. Things have progressed, and. Where. This. Can take us so, they are a point and it's, not that in just one, or, two classes are going to finish this off as we. Will be proceeding. Through the course time. To time we'll, try to go back and see you know where, it all started that. Will kind of give you an idea that where. To go if you do not know the history if. You do not know the time how, it is it has been progressing, it is very tough to build up a story what will be the next landmark. Thing which is going to come up there. So, on. One. Hand. By. When, I started, the lecture today I, told. You that this is used as a technique for certain people yes. There. Are a. Lot. Many, people who have spent their lifetime, in. Or. Devoted. Their lifetime, in. Discovering. Or. Making. This field to, march ahead. So. Keeping. This mind let's. Go. Little bit back to 18th, century. When. The. Cellular. Theory was which was given so one of the theory. Was. Hominid. Cellulite, cellulite, means. Self. Existing. From pre-existing cells. So. Here I will just take a slight, detour for, you. Course. Title is cell culture technology and in this particular next. Series. Of 40. Lectures will be exclusively. Concentrating. On animal, cells this, is just for, your note we, are not taking plant cells into consideration. Here so. That will become way too diverse we'll, talk only about animal. Cells so cell, culture technology you can put it within bracket animal cells okay and of. Course we will be talking about all sorts of animals starting. From mammalian, cells. Even. Human, cell culture to. Mammalian cells. Like. Rat Minds all. The way to amphibians. Like lower. Order. Including. Fishes okay this is the range the spectrum, what we'll be covering, is. Coming back to, the, basic. Fundamentals. Cell. Arises. From pre-existing cells. So. That means. Each. One of each of our cells, in our body arises. From the previous, one if, this is true. Then. Could, we do. This thing outside. The system, what. Does that mean that means say for example I have, these you know epidermal, cells growing. All, over the skin I take, a small part of it, so. What I will be taking out will be a small tissue, so. All of you are aware about the organization, you. Have cells. A. Cluster. Of cells performing. A common function is.
Called A tissue I take. A tissue, so. Each cell is adhering, to the other cells with. The cementing, matrix, I. Dissolve. The cementing matrix, so. What I have are individual, cells and if. I take this cell outside the system, so, no more in part of my body now and plate. It somewhere. Will. It behave the, same way as it happens in my body. It. Making sense let me just put. It now. Graphically. For you to appreciate it okay so. Start. Off with. Your. Cell, culture. Technology. Okay. And we'll. Be dealing as I mentioned. Exclusively. With. Animal. Cells. And. What. I am trying to tell you is that say, for example. From. An animal. I. Isolate. Or. Dissect. Tissue. Now. Once i dissect this tissue. It. Will be something like this. For. Example. The, mass of cells which I have. Isolated. Okay, now. Once. Isolate, this I. Have. Two options either I. Just grow. Them as it is which. We will call as. Outside. The system now up to this this was all inside. Which. Is vivo. So. There is a term which is used for this if you are directly studying, within, the animal is called in vivo now. I have, an in vitro conditions, which is outside the system, now. I directly grow this piece of tissue. Growing. The. Intact. Piece. Of. Tissue. Outside. The. Body. In. That, situation, I will call it a. Tissue. Culture. Of. Course, if this is a dividing, cell then I will be expecting my. Expectation. With this in a tissue culture dish will, be. These. Are the. Four. Cells. Which. Are present. Then. Going. By the theory, of omnis, cellula holiday. Sales. Existing, arriving. From pre-existing cells, i should. Be able to see the. Development. Of the new cells the one which are shown in red so. They will be dividing, and they will be forming a mask kind of structure, right these, red ones are the once, which are. Growing. Outside the system or, what, I do i dissect. This. Tissue and I. Follow. Another route, where. I. Dissociate. The. Tissue. Into. Single. Cell. Suspension. What. Does that mean. It. Means, suspension. Okay that, means. So. I had this. Tissue. Here okay. The. Original tissue which I took. Out from the animal. So. This tissue has, multiple. Cells and each one of these cells are, anchored. To each other where I am putting this small, small red, stumps. These. Are the extracellular matrix. Or the cementing, material, which. Are. Making. These cells. To adhere to each other okay, now. What I do I. Take. Some kind, of an enzyme or some kind of a system by. Which I, break. Down or cue, away those. Cementing, materials so let me introduce that, so. I introduce, some form of a compound. Which will start to nullify them. Okay. So. Once. This cementing, material, is. The. World. What. I will be leaving behind, will, be after. This. Taking. Put this is the. Cell. Cementing. Material, and. I. Am not introducing any technical terminology at, this point will come later on to all those things so cementing. Material. And. So. These, are. Compounds. Dissolving. Cell. Cementing. Material. What. We will be getting after that will be a. Suspension. Single. Cells like this which. Are no more adhere to each other they. Are. Suspension. And this is called the. Word which I used in the previous slide. Single. Cell, suspension.
So. This is called. Single. Cell. Suspension. Now. If this single cell suspension is. Given. A right environment, to grow say for example in a dish or somewhere, and. Okay. Let me introduce this complexity, let me finish this and then I will introduce this complexity. What. Is that and where I stopped, if. I allow them to grow on. A. Synthetic. Environment and, they. Should be able to exactly, behave. Like. They should be able to divide like this so the green is showing they're dividing, which. Is proving the point, omne, cellulite. Cellulite, we, say it. Cells. Arising. From. Three. Existing. Cells. Okay. Now. Second. Situation what we told, you is called. Cell. Culture. The. First one is tissue. Culture, second, one is called cell culture. So. There, will be these. Two words have been used. Very. Haphazardly, in the literature, because, when, Harrison I told you that 1903. The first paperweight, Harrison when. Harrison, did actually, what Harrison there was something like. Tissue. Culture, it took a explant this is that's why it is called next plant it, took a explanted, power of the tissue and grew. It on, a spider net and so, it was I think it was a neural tissue and it shows. The. Extension, of the neural cell board a neural. Cell process is moving. Along. The. Threads of the. Spider. Net okay so. Essentially. The. Term, which is coined was tissue culture. But. Over. Period of time what. We'll be talking mostly about cell culture because the, reason is. There. Are different terms for all these things when. You start with single cells you allow it to grow to. Form a mass or form, a tissue there is the next level if you allow the tissue to go in a three dimension, that's. Called explant sometimes. People directly. Get vex plants so, here, we'll be dealing at different, levels so to start up with in order to sing the course title is cell culture so i told you this is purely what is the cell culture but, we'll be covering the whole spectrum from cell, culture to tissue, culture, to its plant culture, all those things we'll be covering, but, for, the. Basics, it should be very clear to you what, you are doing and. Now. We are into, twentieth, twenty-first century, so we should not use the wrong because, you, know the wrong it's just a confused. So. It is better to use the right terminology what am i doing I am doing cell culture am, i doing tissue culture am i doing explan culture it should be very clear to you there. Should not be any room for any, ambiguity, now. Coming back, if. It is harmony cellulase, allure a cell. Arises. From the pre-existing, cell by rudolf virchow, when. He proposed this which, is part of the, basic. Principles of cell, theory, now. The, question is yes. Indeed. What. Here when I am drawing that, yes from the pre-existing cell, I am getting another cell. Sure. Sorry. I mean out here out, here you, are definitely from, cell this, green one. This. Is the new cell, now. The question, is. How. Much. Closely. This new cell which arose, from the pre-existing cells. Is, similar. To its current so, if this one is the parent, this. Is the parent cell okay this, is the parent cell. And. This. Is the. Progeny. Cells. Yes. They look similar. But. Are they truly similar. In. Terms of. Structure. So. If I had to compare between these two what I will be comparing is. Structure. And. Function. Are. The, same. Not. Only that are their. Structure, function, same. As. Their. In. Vivo. Counterpart. What. Is the enviro counterpart. Here. Are. These cells say for example, if I just add, a little bit more here like. You know. If. I. Talk. About the cell. Cultures here these, individual, cells which are growing, instead. Of the tissue and, if. These are their next. Generation. Forming. Are these. Cells properties, of these cells, these. Cells are. They. Equal to. The properties, of these cells. If. It is so. There. Are two options hypothetically. One. They. All behave exactly the same way the same waving all their, expression, profile of different proteins, all. Other parameters are same or. Being. Outside, the system, the. Course when. They are inside on inside. The system any cell which is going here it. Is exposed to blood vessels it. Is exposed to other tissue it, is exposed to any number of things the. Very moment I take this part or any part of the body and growing it outside in the system it. Is in a totally different environment it's. A totally, synthetic totally, different thing it may, do many, bizarre things witcher. Inside. The body it cannot do say, for example a cell. Which, is growing, here it is under normal control, of several, parameters it, cannot there won't be any outgrowth or anything but, same cells there is a possibility, if i take it out and grow it outside it, may behave like a cancer cell it may behave like something else which. We have no clue because.
It Does not have any restriction, boundaries. It can grow so. How, close, when, we are growing something, outside, the system how, close, we are to, the. In vivo setup, because. That will determine a lot of things that. How what. You are interpreting your experimental, interpretation. Of using a technique based. On this and you wanted to extrapolate, and claim. That this is what is happening in an animal will, depend a whole. Lot. On. How. Close, these, two systems, are and, if they are not close enough, how, far they are so, say for example now let's, draw. The. Lines. Okay. So. What we'll be doing in, the next class we'll start to draw the lines. That how. Close, are, these. Cells to. The there. In. Vivo, counterparts. Okay, so. Let's close in here for the first class and we, will go to the next class where we will be discussing, this. Similarity and dissimilarity and, we'll, come to a, conclusion. That. Where. All the real challenges, lies okay. Thank, you for efficient listening. You.