Vasoactivity Screening

Measure vascular contraction in 3D with high throughput

Our screening service includes:

  • High-throughput screen for compound effects on vasoactivity
  • Real-time contraction data
  • High-content experimentation

Vascular “rings” of A10 rat vascular smooth muscle cells exposed to blebbistatin and norepinephrine

In the drug discovery process, cardiovascular liabilities are one of the main reasons for the failure of a compound. Yet, current compound screens for such liabilities are limited. The current standard is wire myography/aortic ring assay, in which a vessel segment is exposed to a compound and the contractile forces and shape change are recorded. However, this method suffers from the high cost of obtaining animal tissue and low throughput, limiting its widespread use to screen compounds. A need arises for an in vitro alternative to this assay that replicates vascular smooth muscle contractility to screen for these liabilities earlier in the process. Doubly, in vitro alternatives allow for the use human cells to predict in vivo response.

n3D’s solution to this problem is utilizing our magnetic 3D bioprinting system and BiO Assay to print 3D “rings” as a surrogate for wire myography (ex vivo aortic ring assay) to phenotypically screen for vasodilation/vasoconstriction. Not only do these rings resemble blood vessel segments, but when exposed to a drug, they respond similarly by contracting and dilating that we can track in real-time (see video above). Unlike wire myography, this assay is repeatable, high-throughput, and high-content and can use human cells at a significantly lower cost. And unlike other in vitro assays, our aortic ring assay is in a representative 3D environment.

Check out our results presented at ATVB 2014

1. What cell types can I screen with this assay?

  • A10 rat aortic smooth muscle cells
  • Human primary aortic smooth muscle cells

2. How many compounds can I test?
We can screen compounds in multiples of 4, each with 8 concentrations and in triplicate.

3. What is the general protocol?

  1. Aortic smooth muscle cells (ASMC) will be incubated with NanoShuttleTM-PL overnight
  2. ASMCs will be printed into rings the next day for 3-6 h
  3. Rings will contract for at least 5 h, imaged at regular intervals for the length of the experiment
  4. Images batch analyzed to yield contraction data
  5. Post-assay experimentation on rings to investigate mechanisms of action

4. High-content screening is possible with post-assay experiments. What can you perform?
We can do a number of post-assay experiments, including but not limited to:

  • Immunohistochemistry
  • Biochemical assays (MTT, Live/Dead, ELISA)

5. What will I receive?

  • Detailed, concise report of results presented in meaningful way
  • Online presentation and meeting to discuss results
  • Raw data, images, and movie of contraction

Questions? Interested? Contact us for more information and to receive a quote!