A LITTLE MOLD CAN BE A GOOD THING
“[Some] of the problems through the years have been that the formats for 3-D cell culture have been expensive, cumbersome [and] difficult to work with,” says Jeff Morgan, president and chief executive officer of Microtissues in Providence, Rhode Island. However, many scientists have been willing to tolerate these problems in order to use 3-D cultures. Indeed, even Harrison’s original hanging-drop method remains a staple technique in some areas of cell biology, despite its limitations.
Hoping to make 3-D systems more convenient and flexible, Morgan and his colleagues found a surprisingly simple solution: agarose. Cells grown on the ubiquitous gelling reagent can’t stick to its surface. That would be a disaster for conventional 2-D cultures, which depend on surface adhesion to form monolayers, but Microtissues turns nonadhesion into an asset. In the company’s system, cells fall into tiny wells molded into the surface of the agarose, where they form spherical structures.
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Please if you haven’t had the chance already read about the 3D Petri Dish in Nature’s lastest issue. Microtissues was mentioned in the Technology Feature Section in an article called A Better Brew.
“There has been a boom of late in 3D formats, and I think the field is rapidly adopting and critically evaluating these technologies.”
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Dr. Jeff Morgan
Associate Professor of Medical Science and Engineering, Brown University
Recently hailed as one of TIME Magazine’s “Top 10 Medical Breakthroughs,” the bio-artificial ovary is one of the most promising developments in fertility medicine in recent memory. Potential applications offer hope to scores of previously irremediable fertility issues. The Rhode Island Science and Technology Advisory Council is proud to have sponsored the development of this tremendous innovation via a 2009 STAC Collaborative Grant awarded to Dr. Sandra Carson, Professor of Obstetrics and Gynecology at Brown Alpert Medical School, and Jeff Morgan, Associate Professor of Medical Science and Engineering at Brown University.
The story began a few years earlier, when Dr. Carson read about an interesting invention developed by a team at Brown’s Center for Biomedical Engineering led by Dr. Jeff Morgan. What Morgan and his colleagues had devised was a 3-dimensional petri dish. Intrigued, Carson reached out to Morgan.
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Question: We are interested in setting up a hanging-drop cell culture system to grow spheroids of consistent (uniform) size for short-term drug exposure followed by size measurement and determination of the number of viable cells per spheroid. Our preferred cell line is the human colon carcinoma HT29. I came across your 3D Petri Dish® product, which may be good alternative to hanging drop plates. Could you please send information that may be relevant to our requirements?
Answer: The 3D Petri Dish® will work well for your application. HT29 cells form nice spheroids and spheroid size is uniform. Unlike the hanging drop which forms one spheroid per drop, agarose micro-molds made from the 3D Petri Dish® form hundreds of spheroids with one pipetting step. Model 12-256 forms 256 spheroids in one step. Model 24-96 forms 96 spheroids with one pipetting step. Agarose micromolds are placed in conventional multi-well plates for short or long term culture, over 2 weeks. Model 12-256 fits in a 12 well plate, model 24-96 fits in a 24 well plate. Spheroid size is controlled by the number of cells seeded.
The spheroids are easily imaged using a standard inverted microscope so you can document changes in spheroid size with drug treatment. Its easy to add dyes such as WST or MTT to monitor growth/metabolism of the spheroids. The spheroids are formed in the agarose micromolds, so no need to worry about small hanging drops falling or drying out.
Our website has detailed protocols on how to use the 3D Petri Dish® for 3d cell culture and how to harvest spheroids for applications such as western blots or RT PCR.
By the way, the 3D Petri Dish® micro-molds are autoclavable and reusable so you can cast lots micro-molded agarose gels.