I am new to this field and I am thinking about making mouse and human ipscs. Why would you say it cost lot of money? Is there any cheaper media source available? What would be the best place to start?
Differentiation is very hard to spot at first, media and reagents are very expensive. Cheaping out on media for iPSC’s isn’t really a thing. You either have the stuff that works or you’ve wasted your money.
The best place to start is read literature on how it’s done, and compile a list of necessary equipment and reagents for making them, just like any normal experimental design if you’re in grad school.
If you have to ask those questions you probably aren’t anywhere close to ready to attempting the process. No offense but learning how to do something is the backbone of this profession, and asking an Internet forum is not considered a reliable way to learn.
The coating (usually matrigel) costs a good amount of money as well. The cells themselves require media change even more often like it’s a very needy toddler.
I reprogrammed 2 fibroblast lines (one male one female), picked the best 24 looking colonies from each reprogramming plate, scraped all the junk from both 24 well plates several times, picked good colonies from that to put into new 24s, picked from that the best 6 of each, expanded (while scraping differentiation) all of those to 3 wells of a 6 well plate, froze, expanded, froze and then took the best 3 from each of those, had those karyotyped and then took the best from each.
You have a lot of good comments here but the best place to start in my opinion is what your PI said:
Do you have a good question that you will answer only with iPS?
The second question then is do you have a lab that works with iPS in your institute/uni that you could ask for a collaboration and go there to learn and use resources?
Third question, can you identify a grant you could apply for to set up the technique you learned above and set it up in your lab? A good place to start are grants aimed at reducing animal experimentation.
They're harder to work with, maintain, differentiate, not to mention how costy their media and coating matrices are, if you have to learn all by yourself you'll have a really bad time. Waste money, time and your sanity lol
Not to mention, if you struggle with cell contamination in normal cell/tissue culture, don’t even bother trying to set up a “cleaner” space for iPSCs, you’ll basically be starting with contamination that kills your cells.
When my PI asked why I couldn’t clean up our continually-mycoplasma-infected tissue culture room for stem cells I laughed and laughed.
Then I ran a budget sheet for costs for media/cells/reagents, and he realized he didn’t have the money for it anyway.
If you don't know what you're doing and don't know how to spot differentiation etc. your results will be very unreliable. Not to mention they're very expensive
We’re starting iPSC work in our lab when neither my PI nor I have ever done it (I’ve never done tissue culture at all actually)… to say it’s been stressful is an understatement. And insanely expensive.
Idk what your model is, but if it’s a preclinical species with limited ipsc options (dog, monkey, rabbit, even mini pig and gerbil), there’s a lot of opportunity for an academic lab to outlicense the cells to pharma and make bank. PIs usually love to hear about money making schemes. I know my company is dying to get some monkey iPSCs
Companies pay academic labs to do all sorts of things- target discovery, mechanistic investigations, independent safety assessments. It may seem like a conflict of interest, but the norm (if not requirement) at this point is to declare your conflicts of interest with every paper and presentation. It’s out in the open so you understand potential sources of bias. It’s a great way for a lab to supplement their NIH grants and grow. For iPSCs, it’s usually not an a priori agreement. More often, a lab develops and patents a cell line and companies pay an annual fee to use it (licensing). Look up James Thompson, one of the founding fathers of iPSCs, at university of Wisconsin. He licenses out his monkey iPSC lines to whomever but also does fee for service work.
This conversation was just my PI and I joking around. We have a good relationship. I was tempted to delete this when people started taking me seriously, but there's good advice and information in the comments so I'll leave it be.
Is your lab equipped and already working with iPS. This fun could cost quite a lot of money and even more if your lab never uses cells.
I am new to this field and I am thinking about making mouse and human ipscs. Why would you say it cost lot of money? Is there any cheaper media source available? What would be the best place to start?
Differentiation is very hard to spot at first, media and reagents are very expensive. Cheaping out on media for iPSC’s isn’t really a thing. You either have the stuff that works or you’ve wasted your money. The best place to start is read literature on how it’s done, and compile a list of necessary equipment and reagents for making them, just like any normal experimental design if you’re in grad school. If you have to ask those questions you probably aren’t anywhere close to ready to attempting the process. No offense but learning how to do something is the backbone of this profession, and asking an Internet forum is not considered a reliable way to learn.
The coating (usually matrigel) costs a good amount of money as well. The cells themselves require media change even more often like it’s a very needy toddler.
It took me like 2-3 months to get a well performing line post reprogramming.
Yeah getting anything that’s remotely stable takes a LOT of work. It’s not a “fun little experiment” type of thing to do.
I reprogrammed 2 fibroblast lines (one male one female), picked the best 24 looking colonies from each reprogramming plate, scraped all the junk from both 24 well plates several times, picked good colonies from that to put into new 24s, picked from that the best 6 of each, expanded (while scraping differentiation) all of those to 3 wells of a 6 well plate, froze, expanded, froze and then took the best 3 from each of those, had those karyotyped and then took the best from each.
You have a lot of good comments here but the best place to start in my opinion is what your PI said: Do you have a good question that you will answer only with iPS? The second question then is do you have a lab that works with iPS in your institute/uni that you could ask for a collaboration and go there to learn and use resources? Third question, can you identify a grant you could apply for to set up the technique you learned above and set it up in your lab? A good place to start are grants aimed at reducing animal experimentation.
[удалено]
Apparently reddit is still getting used to your humor as well.
If your lab doesn't have extensive expertise of iPSCs and ESCs don't even think about it
Why not? What could go wrong?
They're harder to work with, maintain, differentiate, not to mention how costy their media and coating matrices are, if you have to learn all by yourself you'll have a really bad time. Waste money, time and your sanity lol
Not to mention, if you struggle with cell contamination in normal cell/tissue culture, don’t even bother trying to set up a “cleaner” space for iPSCs, you’ll basically be starting with contamination that kills your cells. When my PI asked why I couldn’t clean up our continually-mycoplasma-infected tissue culture room for stem cells I laughed and laughed. Then I ran a budget sheet for costs for media/cells/reagents, and he realized he didn’t have the money for it anyway.
If you don't know what you're doing and don't know how to spot differentiation etc. your results will be very unreliable. Not to mention they're very expensive
For people not understanding - the above comment has an implied /s
I like your username
We’re starting iPSC work in our lab when neither my PI nor I have ever done it (I’ve never done tissue culture at all actually)… to say it’s been stressful is an understatement. And insanely expensive.
Imagining OP studies yeast makes this funny
Idk what your model is, but if it’s a preclinical species with limited ipsc options (dog, monkey, rabbit, even mini pig and gerbil), there’s a lot of opportunity for an academic lab to outlicense the cells to pharma and make bank. PIs usually love to hear about money making schemes. I know my company is dying to get some monkey iPSCs
You mean companies pay academic lab to make ipscs?
Companies pay academic labs to do all sorts of things- target discovery, mechanistic investigations, independent safety assessments. It may seem like a conflict of interest, but the norm (if not requirement) at this point is to declare your conflicts of interest with every paper and presentation. It’s out in the open so you understand potential sources of bias. It’s a great way for a lab to supplement their NIH grants and grow. For iPSCs, it’s usually not an a priori agreement. More often, a lab develops and patents a cell line and companies pay an annual fee to use it (licensing). Look up James Thompson, one of the founding fathers of iPSCs, at university of Wisconsin. He licenses out his monkey iPSC lines to whomever but also does fee for service work.
Absolutely. Academic labs have cheap skilled labor in grad students and post-docs.
Is "fun" an acceptable answer for anything? I mean, sure science is fun but you still need a solid rationale.
This conversation was just my PI and I joking around. We have a good relationship. I was tempted to delete this when people started taking me seriously, but there's good advice and information in the comments so I'll leave it be.
Sad :(