The centre – which is on Stadium Way in Attercliffe – was targeted on Friday night by burglars who destroyed their gates and stole their van.
They are now appealing on social media for help in tracking down those responsible and replacing their damaged and stolen property.
In a post on the centre’s Facebook page, they said: “Sheffield please can you help us? We are devastated. Please share this post far and wide. We are tired of the attacks on our centre.
“Last night at about 9.40pm the Sheffield Animal Centre was broken into and our Luton van was stolen by two disgusting people. Our first priority was to check all the animals in our care - luckily they are all OK.
“These two men stole our Luton Van, which we use to collect donations, and move medical and animal supplies across Sheffield. This means we have no way of doing that anymore.
“They have rammed our gates which will cost £1000's of pounds to replace - literally. We are so sad and angry about this. We need this van back – DESPERATELY.
“If not we will need to raise funds to get a new one, we are an independent local charity that has to raise all its own running costs. We will shortly be sharing a video of the break in.
“This will be shared with the police too. Do you know these men? Have you seen our van? If so please report this to the Police! Can you loan us a van?
“Can you help get donations to our shops? Can you help us raise money for the gates.
“Sheffield - we need the City to help - ultimately the ones that will suffer are the animals in our care.
“To the people who stole our van, and wrecked our gates...Sheffield is on the look out for you!”
If you have any information about the incident or can help the centre in any way please contact them on 0114 289 8050. The van’s registration number is ML58 XSJ.
You can also report information to the police on 101 or Crimestoppers, anonymously, on 0800 555 111.
To donate to the Sheffield RSPCA’s animal centre appeal visit www.gofundme.com/f/raising-money-to-help-the-sheffield-animal-centre.