93 replies to this topic

[AgeVivo]

Hello,

 

This is both

• a test to do a correctly-sized mouse lifespan test at home (I had done a clearly undersized one with C60 a few years ago)
  -- a matter of smell, time, tools etc
• and a test of wether viagra improves health overall / lifespan
  -- as such things are observed in human data (https://www.studyfin...onger-life-men/) and there are various elements in both rodents and mammals that it improves health in various ways notably possibly some forms of dementia (I will post them some time later)

I will post here (as soon as I am registered on the imminst part I will probably move the thread there, to be in a restricted community) images and statistics of the mouse lifespan test.

[AgeVivo]

First, doing animal experiments at home is forbidden by law in many countries. I am not stating where I do it.

 

The goal here is to take a good care of the animals with things that are reasonably safe (notably taken by millions of persons) and that have reasonably good chances to improve/maintain the health of people, I have already some good experience of animal research and notably lifespan tests in mice in an official lab, so in this very specific case I feel that I am doing something good, but I understand that I do not respect the law in its written format. I do this because I think the benefits for people are worth takeing this avenue.

 

As a consequence, unless something unexpectedly positive happened, the experiment will not be approved by an official committee and will not be published in an scientific article. I count on the presence of adequate persons here for the information to be known to adequate scientists (including people I know) if there results were likely useful for human health.

Edited by AgeVivo, 03 September 2022 - 11:23 AM.

[AgeVivo]

I will start with pictures and ages of the mice, as I had been having them for some weeks and want to use this thread as a diary of what they become.
But before that, a two-month work on my side was to find the equipment - I will talk of the equipment later.

 

I went to various petshops and none of them knew the exact ages of the mice, and they tend to send only a couple of mice at a time: this is not good obviously for a mouse lifespan test where each group should be composed of N=30 (at least 20) mice of known age.

• Side note: these are not mice with a given genetic background, like black 6 mice that research labs buy to Charles River for example. These are "wild" mice, but not so wild : they were imported from reproduction centers that try to preserve a wild background, these reproduction centers sell the mice to pet shops without following precisely the age of each mouse. It is a combo: mice can be received at the petshop at the age of 2 months or 6 months.
• Side note2: some people who have snakes at home buy many mice at petshops, as they are food for snakes

Solution: I crossed two males (white, with red eyes) with six females (of various colors), and will do the lifespan test with the newborns (whose age I will know as I see the births) and prepare the lifespan test with the parents. I plan to start the experiment at a late age (ex: 16 or 18 month of age), so it is an experiment started very long in advance

• This has a risk: what if I change houses for example. Do I kill them? Would I know someone with a snake to give him? Friends who would like to take them? /continuing the experiment?
• This has an advantage: I have the time to prepare the experiment while having my hands on the experiment to appreciate many practical nuances of the at home approach

The two males: 

The 6 females: 

 

These were bought on Juy 16, 2022, as the petshop had just received them and thought they were two months old as they looked to be on the younger range of what they receive. So these mice, that I number 3 and 4 for males and 5 to 10 for females as indicated on the picture (I will recognize them based on the colors, except the makes as they are white and quite indistinguishable; I do not present here mice 1 and 2, that I have bought earlier in order to set up the cages well at home and are therefore older, and will not be part of the experiment - my children play with them, they have special food etc) so these parent mice are likely born mid May 2022

 

 

Of note, each mice costs about 5 to 10 $ so the costs are related to other aspects (cages, food, etc) and it might overall cost me about 1000$ for the whole experiment.

Edited by AgeVivo, 03 September 2022 - 12:26 PM.

[AgeVivo]

^ precisions: i) petshops "tend to sell (not send) only a couple of mice at a time" ii) the female mices have black eyes

[AgeVivo]

Mating

On July 19, I put 3 females mice (numbered 6, 9 and 7) with a male (3 or 4, undistinguishable), and the 3 other females mice (number 5, 8 and 10) with another male (3 or 4, undistinguishable).

After a few minutes, the males started willing to mate, the females got afraid and angry and fought, but after a few days the situation was established. On male seemed to be friend with females without trying anything, the other male seemed to be more dominant with alternances between attempts and calm, and in both cases there was no sign of bad reactions anymore.

 

On August 12, I removed the males from the females and put the two males together. Here are the pictures of the females on August 12

 

After only a minute together, the males started to fight with each other. I had a very hard time to put them together - I split them in different environments, provided them with various toys, put them together for a few seconds in a very enriched environment, separated them again, etc. In the evening - a few hours later, things seemed fine although one had a little blood on the side. During the night I could here much noise, hopefully because of the many games, but the next morning one of the two males seemed really in bad mood. The next day it was found dead. This is horrible but I present things as they are.

 

In August 28, as I indicate in the next thread, one female had 8 babies.

Edited by AgeVivo, 03 September 2022 - 01:59 PM.

[AgeVivo]

^typo: 9 babies

born probably on August 26 or 27, based on the non-pink color of their skin (picture on August 29):

I am not used to births, the exact day of birth might be adjusted by one day later based on pictures

let us save the numbers 11 to 19 for them

They are from mouse 6 (black and white). Here is also a picture of the male (white):

[AgeVivo]

One week later, 9 other babies are born, from mouse 7 or 9 (I will confirm very soon; I need to go see what number correspond to which)

Let us save the numbers 20 to 28 for them.

Here are the 18 babies at September 3: 

The difference is very clear between

• those who were born slightly more than a week ago: they have (black) fur, they start to walk and crawl around
• those who were born extremely recently: they have a flask, sticky pink skin, their eyes are somewhat under the skin

Here are also the various adult females, just to keep track of them:

6, 7, 9:  and 5,6,10: 

 

Of note, these 18 newborns are in the same cage as the adult females 6 7 9 (who take good care of the 18)

Edited by AgeVivo, 03 September 2022 - 10:01 PM.

[AgeVivo]

The reason why I take these careful images is to know quite precisely the age of the mice and who they parents (mothers) are, for later survival statistics. Very likely, a birthdate precision of one week is sufficient for the overall analysis and very likely it does not really matter who the parents were, but I prefer to err on the side of caution.

I will also take pictures when the young mice start to be distinguishable by they fur, to identify and follow them well all along their lives.

 

N=18 is not at all enough, but I need to organize the cages and air flow well.

As I will describe (in next post, likely), it is easy at home to have two cages of 5 mice (ex: in the kitchen or in a room with water), but above two cages air ventilation starts to be a matter of attention if cages are not changed more often than every week

Edited by AgeVivo, 03 September 2022 - 10:12 PM.

[AgeVivo]

the newborns 20-28 are from mouse 7

One week later, 9 other babies are born, from mouse 7 or 9 (I will confirm very soon; I need to go see what number correspond to which)

Let us save the numbers 20 to 28 for them.

Here are the 18 babies at September 3:  2022-09-03_11-28.jpeg

The difference is very clear between

• those who were born slightly more than a week ago: they have (black) fur, they start to walk and crawl around
• those who were born extremely recently: they have a flask, sticky pink skin, their eyes are somewhat under the skin

Here are also the various adult females, just to keep track of them:

6, 7, 9:  2022-09-03_6,7,9.jpeg and 5,6,10:  2022-09-03_5,8,10.jpeg

 

Of note, these 18 newborns are in the same cage as the adult females 6 7 9 (who take good care of the 18)

 

[AgeVivo]

A few pages on LongeCity that can help choose experiments to do:

https://www.longecit...ew/#entry918084

https://www.longecit...ad/#entry882022

https://www.longecit...et/#entry917245

[AgeVivo]

On september 11, I moved the mice to cages with a paper litter. I will do this from time to time as traditional litter, wood-or-similar- based, can irritate lungs, so this helps lungs recover from irritations.

 

Female mice 6,7,9 (none of them had babies in the summer): 

Female mice 5,8,10 (2 of them had babies): 

The babies 11 to 28 (actually, there was not 19 but 18 babies; I check in very fine details all over the cage and did not find any trace of a baby that could have died / been eaten, so I likely doublecounted one of them last time, and indeed it wasn't easy to count them; one baby is more active than others as we can see in the picture and I had already seen this the week before so it is probably the same one, but all of them have black hair - I hope hair color will change later in order to be able to distinguish them...): 

I have then put females 5 8 10 together with the babies of course

The male, numbered 4: 

I have them to put water, food and toys back, of course.

[AgeVivo]

This Saturday I changed the cages of the cages except for the adult male as it is alone in its cage that is still clean

Female mice 6,7,9 : 

Children 11-27 (9 older, 8 younger; all of them are black; I hope it will change, to be able to distinguish them) : 

With their mothers and the helper (3 "mothers") and lots of toys (now that they start to be able to eat, drink water, climb and jump, I added a wheel and made a large playground, it changes the atmostphere from a nursery to a playground for children monitored by parents, they seem to like it much) 

 

The road to a life extension test is still very long ahead however... Ideally many citizens like me would do this, to screen the most promising protective aspects for health and longevity, at an approximate overall cost of 1000 € and mouse care for 3 years. But I guess this will not happen.

I asked for a quote to a company, such a lifespan test in standard lab conditions costs more than 50k€... I guess that financing many mouse studies outside of homes won't happen either ; not more than what we see in the scientific litterature.

 

Still, I believe this is one of the keys to our future health and longevity.

I recently read for example that statins were demonstrated with 24-year long studies in humans to increase cancer risks (contrary to what most think, having somewhat high cholesterol is cancer-protective and not linked with cardiovascular risks, and lowering it with statins creates cancers over time), something known with mice and rats since the 80s, but commercial interests on statins tried to conclude the contrary and more than 20 years were needed for truth to be established (not that some statins are some doses are not overly beneficials, such as Spindler's simvastatin study in mice, but then the effects are slim). All this to say, the integrated mouse lifespan model remains a needed model for life extension

 

So.. let's continue this experiment that I here blog about, for the next 2 or 3 years to come...

Edited by AgeVivo, 17 September 2022 - 04:40 PM.

[AgeVivo]

Cages changed again

[AgeVivo]

Cages changed again,
The three adult females 5 8 10 who did not have babies are of course still together : 

 

I sadly see that I am sadly not able to distinguish between the two generations of newborns ; the ones born at the end of August versus ones at the beginning of September. This is because 1) all of them are essentially black. I was not expecting this as coming from a white father (with red eyes - "black" males with recessive albino genes ??) and largely white mothers Had I separated the two generations immediately, I would still have avoided this issue. More rapidly than I thought, it was too late to see who was young.

So it will be an issue for my experiment. I will hardly recognize individual mice based on their colors and I will not know age more precisely than by a week.

 

I put the (2+1) mothers and (10) daughters together:  (sorry for the ugly picture, I put them in that bad cage for a few minutes, time to have their clean and fun cage prepared)

 

And of course I have separated the (7) sons as they start to be adults and I don't want to have a million mice: 

 

Filling OK to handle childhood now, I have put the father together with another colored mouse (that you have not seen yet, it is the mascot of the family as it was the first bought and is probably one or two month older than the other 6 clearly-adult females. To limit the risk of fight, I first separated them in the same cage:  , then let the female decide to go to the other border and leave when she wants and vice versa, and then put them together by removing the separation. The male is willing to mate, the female not really, but it doesn't seem to fight. I put a thin cup on the side to that the female can protect herself. I hope they will get along each other, and will separate them otherwise.

 

Here are all the mice, in clean and fun cages: 

Forefront: sons on the left (they start to have mini fights as they get teenagers/adults, but nothing bad), females on the right (mothers + daughter; fun and peaceful cage)

Back: females on the left (zen cage, they like their round-shaped house and the discrete entrance), new couple on the right (the two mice are happy not to be single and to be together but they do not agree about mating)

All of they know me/my hands and are happy with me tweaking their cages.

 

Once the couple will have babies, I will need to have at least two more cages and to switch to a different installation in terms of air filtering. I have several ways to do it but need to think about it and do it within the coming month. It is not easy to have enough mice for a lifespan experiment at home.

[AgeVivo]

cages changed. didn't take pictures but all are healthy, nothing special to declare

[AgeVivo]

a bit after I wrote the previous message, I observed that one of the three mums was pregnant  as it was wider than normal (mother 7; as a reminder, mother 6 had babies at the end of August and mother 7 early September). So it means that before I separated the sons one of them made mother 7 pregnant. It must have been one of the oldest ones so from mother 6, so luckily it is not a bad genetic mix.

 

This is actually good because I needed more mice for the experiment and female 1 and male 4 don't seem to be able to have babies (I wish them had; male 4 is the one who could not create babies with females 5 8 10, and who killed the one who created babies with females 6 and 7...) 

 

Yesterday female 7 was very pregnant, so I put the three "mums" on a separate cage (2 biological mums in fact, but these three females were in the same cage in the petshop and have always been in the same cage later, and have always helped each other) from their 10 daughters (the 7 sons were already on a separate cage) and she had her babies this morning.

13 babies I think (it could be 14; or 12; I counted visually in order not to disturb). All pink and about 1 cm long.

 

So here is the troup:

The 3 mothers (6,7,9) and their 10 essentially-black daughters yesterday (by deduction the father was a black albino), just before I separated the 3 mothers:

The 7 essentially-black sons of the 3 mothers: 

What the approx 13 newborns from today look like (after I watched them, the mother burried them): 

 

The 3 non-mothers (5;8; 10): 

 

The non-father (4) and the first female (1):  As you can see I have tried this hemp litter ; here is doesn't seem great but to my surprise they really love it (they put themselves in the fold, they made two wholes in it to hide themselves, it is like a bed cover to them; as a reminder, wood chips should be avoided as it damages lung. So next weeks I will test the hemp litter in other cages. 

 

Given that I need more than 4 cages I have split the aspiration tube as we see here (using scisors and a bit of electrical tape, not as complex as it could seem):  I am limited in surface by 3x2 = 6 cages.

 

For now I use 5 cages for a total of (3+10+7+13)+(3+2) = 38 mice, including about (3+10+0+6)+(3+1)=23 females and 15 males. I think I can go up to 9 cages and 80 mice maximum in this area, I think that the air aspiration is strong enough not to feel uncomfortable due to smell. I go very gradually there as I need to learn how to master all this so that it doesn't take too much time, doesn't smell bad, so that I don't lack water, food, have no mouse escape etc. I think that in 1 month I will have:

Cage1 - 10 daughters 1

Cage2 - 7 sons 1

Cage3 - approx 6 daughters2 and 3 "mums"

Cage4 - approx 7 sons2

Cage5 - 4 females and one male (if the merge works well, otherwise 2 cages)

Although many things could happen.

[AgeVivo]

Oops, previous post I used an old image for mice 5, 8, 10. Here is today's image: 

 

Here is an overview of the installation (the picture shows extreme light and as a resut superimposes the two rows of cages, not sure why; in practice the light is approximately the same everywhere): 

Edited by AgeVivo, 23 October 2022 - 03:21 PM.

[AgeVivo]

The 12 newborns (yes, there are 12, I wasn't sure in the last post as I was counting them from far)
appear to have a diversity of colors. This is what I was hoping (also for the first series of newborns...)

We see them here with their biological mother (#7), their grand-mother and possibly aunt (#6),
and another possible aunt (#9). The three "mothers" (who were bought together from a crowded cage at a petshop)
take approximately equally care of the newborns, just as they did for the first series.


The father is one of these 7 males, from the first series of newborns:

Here are the 10 females, from the first series of newborns:
Interestingly, 4 of them are gray.

Here are the three other friends/sisters of females 6 7 9:
I did not take in picture the couple that will not have babies it seems

Here is the installation with all the mice:
Behaviors:
I modify the organization of cages every week because it keeps them extremely curious and makes them happy.
They seem to like these houses very much; and this hemp litter very much because they can fold it in tunnels
and pull fine fibers from extremities to make comfortable beds and block the entrance of houses.
They prefer to run in the wheel but exceptionally I moved some wheel out of its support for them to investigate.
Adult mice start to all like me and want to crawl on my hand and arm, I avoid it for them not to get lost (nor fall one day!).
Among the males of the first generation, one of them was byting the ass of others and being quite mean at first
(sort of young-teenager behavior) and gradually it has transformed as a nice game, which other males now like.
Apart from this, they respeci each other very much.
Males want a bit more of challenge than females, so I have invented various challenges,
the one in the air on the top right is their preferred one (a suspended platform that flips when climbing on bars;
they hand on the platofm and bars in order not to fall and try again).
They gradually coordinate across each other inside wheels, to do loops.

Attached Thumbnails

• 

[AgeVivo]

Good, the babies can be distinguished quite easily:  : 2 black and white , 4 gray and white , 2 fully gray , 3 black

 

Here are the 3 mothers , their 3 sisters:  ,

 

the 10 males (incl 4 gray) and 7 females of the August/September cohort 

 

and the couple that does not have babies (that I call "independant", due to their quite independant caracter) 

 

I have noted that with standard hemp litter I put 2.8L per cage. This, to soon estimate the cost of mice. I had the information from someone working at the ITP that getting a 18 month old mouse costs about 100$, I will try to compare.

Edited by AgeVivo, 06 November 2022 - 12:58 AM.

[AgeVivo]

1. I was surprised to count one newborn less than the week before, just like with the late August/early September cohort at the same age. I googled and it is well known mice eat some of their most immature children around age 10 days when not able to handle them all, and this particularly happens when one changes the places of the newborns. I was told about it in my past when doing animal research but it is clear. I guess I have to accept it (or not clean cages at that time)

2. with 100L of standard hemp litter (link), one does about 35 cages so an estimation of the cost of such litter per cage and week is 27-to-30$/35 = 80 cents (perhaps 85).

with 25 m of carpet hemp litter (link) at about 70$, one does 75 to 100 cages so an estimation of the cost of such litter per cage and week is 70/75 = 70 to 100 cents

so I was wrong about the difference of cost between the two. It depends on how much one put in a cage.

 

For a cage of 6 mice (one gender type out of 12 newborns) of 18 months (78 weeks), taking a cost of 1$ per week (adding costs to clean cages: paper, water...), considering the same for food (this is probably less, here I am really not precise yet) and adding 100$ initially per cage (cost of cage and its materials), we end up at (78*2+100)/6 = 43$ to have an old mouse. This is of course without counting my efforts every week-end The 84$ at the ITP is very cheap (they cover perfect standards, with adequate pressurizations, trained personal etc). Though, when I asked companies to do a mouse lifespan test with a compound that I provide it would cost 60k$ with N=30 per group, which I don't easily have, and in my case if I take 30 mice per group, and count twice as much as what I counted for reaching 43$, in order to take the compound into account, it costs 43$*2*60 : about 5k$ -- 5 times more than I initially thought at the beginning of this thread; perhaps I took too high estimation margins here.

So the biohacking experiment is "only" 12 times less costly than the recognized experiment. Is it worth it? Not obvious (no official publication, very borderline) but when one does this at home in week-ends aside from job and family (because biology research does not pay well and one has to pay the bills) and does not have 60k$ at hand, it is about the only solution. The other one is crowndfundings with researchers and the longevity community.

Edited by AgeVivo, 06 November 2022 - 10:57 AM.

[AgeVivo]

Changed the mice; no time now to post all pictures so in a nutshel no change in configuration since last week, all is good and simple

 

The number of interventions I would like to test is of course large. I am thinking about making a big mouse center in my garage; techically given the speed at which mice make children and if one is able to spend a full day every week end (except exceptions during vacations) it seems at hand. Though I am not in an isolated house so it gets tough. Perhaps someone is better positionned than me for this ? Ideally research grants would organize the MMTP but it is not quite the case so far. All are isolated tests, that don't provide that much for longevity

[AgeVivo]

How to be remunerated to build a mouse facility that performs massive longevity tests ? The reason why I do things at home is that I didn't find - the few times when I tried it didn't go till success, and my time is limited to try again and again. To pay the bills, for now I need my basic work. But would someone come with a clear, established funding, of course I would jump for it

Edited by AgeVivo, 13 November 2022 - 07:40 PM.

[AgeVivo]

Changed the mice,

 

The while male... killed the female he was with. Third mice he killed - he is actually the only mouse of mine that has killed others (except very young newborns by mothers, something well documented). So a serial killer. And now that he is alone, he doesn't seem happy and is waiting for me to move things around. I did not take him in picture

 

The six sisters, of the same age probably as the white male (all were bought), including 2 that gave birth to mice, are here:

 

Their 10 females are here: 

and their 7 males are here:  (one likes to bite others and they are afraid of him... no death so far but not good for health and lifespan I guess... I wonder if I should move him away, put him with the white male but it may lead to another death.. Not obvious choice - If one had experience on such things, do not hesitae)

 

Here are the 11 children, from one of the 6 sisters and one of the 7 males: . I will soon need to split males from females

 

And here is the overall installation: 
Thanks to the aspiration is doesn't smell - I have reached a very stable installation, with happy mice - except for the two strange males - and I change the cages in a few hours every week-end 

[AgeVivo]

Separated the boys and girls in the second set of offsprings, as they are one month old (slightly more). A bit disapointing: only 4 females (on the bottom of the picture here) for 7 males (on the top), whereas I aim to do a lifespan test in females

 

Regarding the first set of offsprings, here are the 11 females and 7 males (+the white male, that surprisingly gets well along with them; out of the black males one is bitten on the back, but I knew this already as one of the males regularly chases the other ones and bites them)

 

Regarding the 6 sisters initially bought, there was a big fight two days ago and one died. I feel bad that each time I put groups together (even the 6 sisters that were separated for a few weeks), things get dramatic. The lesson is that now that I am able to handle things well, I need to create a cage with many newborns and split them, rather than have too small groups that I need to assemble in order not to have too many cages. But so far I could not do it, so no regret. Here is also the general installation

 

[AgeVivo]

Plan of experiment:

I am in the situation on the left, if I buy 3 males and that 3 females become pregnant, I should have the minimum number of animales needed who are of known age (that is, not bought but born at home):

[AgeVivo]

Plan of experiment revised:

It is unlikely that out of 3 females all of them become pregnant, and the 2 ones that were already mothers can be again, I see no issue with that (these are not albinos mice, that were bought, there is not reason they have strange/non-representative-to-other-mice genetic features). So I will put all 5 bought females with bought males; and I probably need to buy only 2 males. Will do so tomorrow.

We will see if I get about 20, 30, 40 or 50 newborns: I see that I am still learning how to handle these things with mice.

[AgeVivo]

I did the latter, second phase (no babies yet), the white male was killed by the seven black males (not surprisingly I must say; at least now I know); it took a week for the 2 males to live correctly with the 5 females, for now all is good but something could happend. Apart from this very high territoriality of mice, all good, mice seem healthy and happy.

[AgeVivo]

4 females 
+ 10 females 

+ 7 males 

+ 7 males 

 

+.. 2 males and 5 females hopefully preparing the complement for the lifespan study

[AgeVivo]

All is fine in all cages ; except that in males cages there is one who likes byting others to be the chief, but that is normal mouse life... ; and that in the cage with 2 males and 5 females, no baby for now

[AgeVivo]

It has been a long time since I haven't posted.

I currently have 53 mice in 5 cages, and I will very soon have enough to stop letting babies be created and just focus on an lifespan experiment:

• 29 females
  • 5 colored females aged 8.1 months (bought together - "sisters"; currently all of them in one cage, "3a", with two bought males, two "second offspring" males; although one "male" migth be a female, I'll check next week-end)
    
  • 10 black females aged 4.5 months ("first offspring", cage "4a") 
  • 4 colored females aged 2.8 months ("second offspring", cage "1a") 
  • 10 females (estimation, I'll check next week-end) aged 3 weeks born from the latter one ("third offspring", cage "1a") see "23 babies" here below.
• 24 males
  • 7 black males aged 4.5 months ("first offspring", cage "2a") 
  • 2 colored males aged 2.8 months (second offspring, cage "1a" seen here above)
  • 2 bought colored makes aged 2.8 months (cage "1a" seen here above)
  • 13 males (estimation, I'll check next week-end) aged 3 weeks (separated today from the females: moved from cage 1a to a new one: 5a) 

 

Cages 1a and now 5a correspond too 23 babies born on December 23, 2022, from 2 mothers (of the 2nd offspring) and surveiled by them and 2 other females. Here are pictures prior to the ones above.

• As they were born, there were 24 babies but the two mothers started eating one of them. I immediately took it, they ran after it, I gave it back and they hurried up to eat it. It must be a physiological process because they had plenty of food and water around and there was a nice atmosphere in the cage. 
• During the first week, then were crawling together in a transparent dome. In the second week, some started to try to go out but the mothers made them come back. When changing chages, I put the mothers on the side then move the children to the new cage and then put the mothers back, and here the mothers started letting the children go out. 
• 3rd week, this is the last picture of all of them together just before putting males aside, as males start to show interest for females and I am reaching the right N . I will double check next week-end if I did the split between males and females right, because at age 4 weeks it is much more visible to me than at age 3 weeks