good evening and welcome you all to this new course which will be starting today on cell

culture technology so the name cell culture whenever it comes gross people have the first

feeling that well its a lab training or a kind of a technique which is used by most

of the labs who worked at the interface area biology chemistry environmental sciences and

all other allied fields might metabolic sciences say for example so its just a simple technique

what most of the kind of understand that you know cells which are cultured in an synthetic

or artificial environment at least thats what the commoner or ah you ask any graduated student

or ah under graduate student thats what they will answer why we really want to offer a

course on this area

so and as a matter of fact when i floated this course there are people has been that

thats a practical training how really want to you know put a cross a course which is

mostly a practical training people have cell culture labs so for me to 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 ah 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 mile stones then it will be

easy for that individual to design the problems in 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 logics behind the spectroscopy whether

its a raman spectroscopy whether its a ir whether it is a nmr if you know those at least

the basics then you know with your problem what all you can derive using this spectroscopy

or say for example crystallography similarly several mathematical tools similarly

like a statics 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

our problem similarly cell culture is a tool but tool whose journey if you look back is

now more than hundred years old and there are handful of books you will come across

and some of them i have already mentioned like m tissue cultured book or ah few other

books like gorsline bankers neural cell culture book and there are manuals published by companies

like gibco invitrogenor currently which is known as 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

has such as not just a mere protocol is definitely there is a significant part of it which is

protocol driven just like an molecular biology in a manual even you can go through it ok

but a still molecular biology has a subject has tremendous amount of chemical logics and

unless you understand that you will be working like a blind person thats precisely what happens

in cell culture that my young student or young enthusiast take under graduate or a post graduate

just embraces that this is technically just let follow it

without putting logics and place that why am i doing it what is the basic reason behind

it so in this course what we will do 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 ah if you look back like if

you are an 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 were around

nineteen o eight nineteen hundred and twelve it is that back even much earlier than the

first world war first world war started in nineteen fourteen right

so this was the paper from harrison one of the pioneer and what he did for the first

time he grew a explant explant means part of a some animal systems body or a matrix

and a matrix was very interesting thing it was a spider net he didnt know where to grow

so if we break up this problem what he was trying to do but before what is trying to

do lets go back a little so if this field which has its first set of publication coming

almost hundred seventeen years back then for most what we will try to do we will try to

appreciate this whole history of hundred years what is happened how things are progressed

and where this can take us so there were points and its not that in just one or two classes

we are going to finish this off as we will be proceeding through the course time to time

we will try to go back and see you know where it all started that will kind of give me 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

land mark thing which is going to come up there

so on one hand as i when i started the lecture today i told you that this is used as a technique

for certain people yet there are a lot many people who have spend their life time in or

devoted their life time in discovering or making this field to march ahead so keeping

this mind lets go little back to eighteen century when the cellular theory was which

was given so one of the theory was how many cellula e cellula means cell existing form

preexisting cells here i will just take a slight for you course

title is cell culture technology and in this particular next series of ah forty lectures

we will be exclusively concentrating on animal cells ok this is just for your note we are

not taking plant cells into consideration here because that will become way too diverse

we will talk only about animal cells so cell culture technology you can put it within bracket

animal cells ok and of course we will be talking about all sorts of animals we starting from

myelin cells even human cell culture to mammalian cells like rats mice all the way to amphibians

like lower order including fishes ok this is the range the spectrum what will be

covering ok coming back to the basic fundamentals cell arises from preexisting cells so that

means each one 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 is that mean that means say for example

at 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 ah cluster of cells performing

a common function its called a tissue i take a tissue so each cell is adhering to the other

cell with a 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 body now and

plate it somewhere will it behave the same way as it happens in my body is it making

sense let me just put it now graphically for you to appreciate it ok

so we start off with so our cell culture technology ok and we will 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 a tissue now once i dissect the tissue so

it will be something like this now for example its a mass of cells which i have isolated

ok now once i 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 in vivo

so there is a term which is used for this if you directly studying within the animal

is called in vivo now i have in a 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 cultured dish will be these are the cells which are present

then going by the theory of omni cellula e cellula cells existing arriving from preexisting

cells i should be able to see the development of the new cells the one which i have shown

in red so there will be dividing and there will be forming a mass kind of a structure

right this red ones are the ones which are growing outside the system all what i do i

dissect this tissue and i follow another route where i dissociate the tissue into single

cell suspension what does that means let me suspension ok

that means so i have this tissue here ok the original tissue which i took out from the

animal so this tissue as multiple cells and each one of the cells are anchor to each other

where i am putting this small small red stumps these are the extra cellular matrix or the

cementing material which are making these cells to adhere to each other ok now what

i do i take some kind of an enzyme or some kind of a system by which i break down or

chew away those cementing materials so let me introduce that

so i introduce some form of a compound which will start to nullify them ok so once this

cementing material is dissolved what i will be leaving behind will be after this ah let

me put this is the cell cementing material and i am not introduce any technical terminology

at this point we will come later on to all those thing cell cementing material and so

these are compounds dissolving cell cementing

material what we will be getting after that will be 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 day short somewhere and ok let me introduce

this complexity let me finish this then i will introduce this complexity ah 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 this is green

is showing the dividing which is proving the point omnis cellula e cellula cell arising

from pre existing cells ok now second situation what i 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 ah very oppositely in the literature

because when harrison i told you that nineteen o three the first paper by harrison when harrison

did actually what harrison did was something like a tissue culture he took a explants this

is thats why it is called an explants he took a explants or part of the tissue and grew

it on a spider net so it was i think it was a neural tissue and it shows the extension

of the neural cell bar neural cell process is moving along the threads of the ah spider

net ok so essentially the term which is coined was tissue culture but over period of time

what we will be talking mostly about cell culture because the reason is there are different

terms for all this things when you start with single cell you allow it to grow to form a

mass or form a tissue there is a next level if you allow the tissue to in a three dimension

thats called a plant sometime people directly get the explants so here we will be dealing

at deferent level so to start of with in order to since course

title is cell culture i told you this is purely what is the cell culture but we will be covering

the whole spectrum from cell culture to tissue culture to its plant culture all those things

we will be covering but for the basics it should be very clear to you what you are doing

and now we are into twenty first century so you should not use the wrong because its not

the wrong its just the confused so it is better to use the right terminology what i am doing

i am doing cell culture am i doing tissue culture am i doing explants culture it should

be very clear to you there should not be any room for any ambiguity

now coming back if it is omni cellula e cellula cell arises from the pre existing cell by

when he propose this it just part of the basic principles of cell theory now the question

is yes indeed what here i am drawing that yes from the pr existing cell i am getting

another cell sure sorry i mean out here out here yeah definitely from cell this green

one this is t new cell now the question is how much closely this new cell which arose

from the pre existing cell is similar to its parent

so if this one is the parent this is the parent cell ok this is the parent cell and this is

the progeny cell yes they look similar but are they truly similar in terms of structure

so if i have to compare between these two what i will be comparing is structure and

function are they same not only that are their structure function same as their in vivo counterpart

what is the in vivo counterpart here are these cells say for example if i just add little

bit more here like you know if i talk about the cell culture here if these individual

cells which are growing instead of the tissue and if these are there 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 behave the same way means all their expression profile

of different proteins all other parameters are same or being outside the system because

when they are inside the inside the system any cell which is growing here it is exposed

to blood vessels it is exposed top other tissue it is exposed to n number of things but the

very moment i take this part or any part of the body and growing it outside the system

it is in a totally different environment its in totally synthetic totally different thing

it may do many blizzard things which inside the body it cannot do

say for example a cell which is growing here it is under normal control of several parameters

it can their own be any out growth or anything but same cell there is a possibility if i

take it 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 set up because that will determine a lot of things that how what you are interpreting

you experimental interpretation of using a technique based on this and you wanted to

extra polite and claim that this is what is happening in an animal will depend ah whole

lot on how close these two systems are if they are not close enough how far they are

so say for example now lets draw the lines ok so what will be doing in the next class

we will start to draw the lines that how close are these cells to the their in vivo counter

parts ok so lets close in here for the first class

and we will go to the next where we will be discussing this similarity and this similarity

and we will come to a conclusion that where all the real challenges lies ok

thank you for a patience listening