ARDF Annual Open Grant Program

ARDF's Annual Open grant program was established to fund research projects that develop alternative methods to advance science and replace or reduce animal use. Proposals are welcome from any nonprofit educational or research institution worldwide, although there is a preference for U.S. applications in order to more quickly advance alternatives here.

Expert reviewers evaluate proposals based on scientific merit and feasibility, and the potential to reduce or replace the use of animals in the near future. Proposals are considered in fields of research, testing, or education. The maximum grant is $40,000, with an average 21.5% funding rate from 2015 to 2020.

Since 1993, ARDF has provided over $3.75 million in funds for projects in 29 states and 5 countries.

Scientific reviews are conducted under the guidance of the Institute for In Vitro Sciences, Gaithersburg, Maryland.

  • Maximum Award per Project: $40,000
  • Open Date: JANUARY 20, 2021
  • Deadline Date: APRIL 30, 2021 
  • Notification of Recipients: JULY 19, 2021
  • Non-profit, educational, and/or research institutions
  • May not use intact, non-human vertebrate or invertebrate animals
  • Research, testing, and education fields
  • Are from U.S. institutions or organizations
  • Utilize in silico and in vitro methods with human cells or tissues
  • Use pathway-based approaches as exemplified by the 2007 National Academy of Sciences report, Toxicity Testing in the Twenty-first Century: A Vision and A Strategy

2020 Grants Awarded

ARDF is pleased to announce the recipients of the 2020 Annual Open Research Grants

Congratulations to awardees:

Departments of Orthopaedic Surgery, Pediatrics, & Bioengineering, University of Pennsylvania, Philadelphia, PA*
3DEndOs - An in vitro toolset to study endochondral ossification

*Co-Primary Investigator Dr. Gottardi’s lab is at the Children’s Hospital of Philadelphia Research Institute
Naturally, bone heals via endochondral ossification, a bone formation process in which progenitor cells first differentiate into chondrocytes to form a non-vascularized, hypoxic cartilage template that is then replaced by bone tissue via matrix degradation and blood vessel invasion. It is well known that bones adapt to mechanical forces, but endochondral bone healing is orders of magnitude more sensitive to mechanical stimuli. However, we know little about how cartilage template hypoxia regulates blood vessel recruitment and mechanotransduction during endochondral ossification. Filling this gap will be crucial to develop new therapies to accelerate bone healing. Current preclinical approaches to study endochondral ossification rely on in vivo model systems that do not enable precise control of oxygen tension or mechanical stimuli, limiting mechanistic understanding. The goal of this project is to use bioreactor models uniquely established by this collaborative team, to define the role of hypoxia-induced signaling in directing vascularization and mechanical regulation of endochondral ossification. We will employ sophisticated bioreactor systems featuring biphasic microphysiological control and mechanostimulation to engineer human stem cell-based endochondral tissues. While in vitro modeling of endochondral ossification has been challenging for the field, our innovative approach integrates three critical components: hypoxia, angiogenesis, and mechanical loading, which we anticipate will be transformative. Solving this problem will enable us to uncover new fundamental mechanisms and develop therapeutic approaches without the need for animal research.

Departments of Medicine and Pharmacology, Division of Endocrinology and Metabolism, University of Washington, Seattle, WA
Development of an in-vitro assay teratoma assay using microfluidic chip technologies
Advances in stem cell research offer new hopes for patients with diabetes, Parkinson’s disease, heart failure, and other degenerative diseases. Yet, for these cells to reproducibly generate tissues of interest for translational use, it is critically important to first assess their pluripotency. The simplest method used is their aggregation and differentiation into 3D clusters called embryoid bodies (EBs) to determine if they can generate all three germ cell derivatives (ectoderm, endoderm, and mesoderm). However, this assay cannot accurately predict the developmental potential of stem cells to generate different tissues. These limitations have led to the development of the so-called Teratoma assay, a test in which immunodeficient mice are injected with stem cells to generate Teratomas, a form of tumors containing cell types derived from all three germ layers. Unfortunately, this assay requires a large number of mice, is expensive and labor-intensive, and has limited reproducibility. In this project we will develop a Microfluidic In-vitro Teratoma Assay (MITA) that overcomes the limitations of EBs and can eventually replace animal testing. Based on our pilot experiments, we anticipate that our assay will not only replace animal testing, but also provide a tool for assessing the tumorigenicity of stem cell-derived preparations, and testing of new therapeutics.

Cancer Epigenetics Program, Fox Chase Cancer Center, Philadelphia, PA
Long-term in vitro co-culture model for maintenance and screening of human primary AML
Acute myeloid leukemias (AML) is the most lethal of all blood cancers, killing three out of four patients within five years ( Despite many advances in cancer therapy, most AML patients will eventually succumb to therapy-resistant disease. This is a critical area in which significant breakthroughs must be made. The discovery of frequent mutations in epigenetic modifiers in AML has led to an intense interest in the development of novel epigenetic therapies. However, epigenetic agents typically require longer duration of treatment to achieve their therapeutic effects and require models reflecting the heterogeneity of AMLs beyond cell lines. This limits the options of pre-clinical screening mainly to patient-derived xenograft models (PDXs) in immunocompromised mice, which are time-consuming, expensive, and involve animals. As an alternative, I recently developed an in vitro 2D co-culture platform that enables screening of primary AMLs with clinically relevant drug regimens without the need for animals (Duy et al. 2019, Cancer Discovery). Yet, the 2D platform utilizes FBS and this proposal will explore serum-free conditions to completely replace animal products. Furthermore, the proposal will establish a novel 3D culture model to assess self-renewal potential of human primary AML in vitro that will replace additional animal models.

Environmental Health Sciences, School of Public Health at the University of Michigan, Ann Arbor, MI
Discovering host factor inhibitors in silico for SARS-CoV-2 entry and replication
Our long-range goal is to advance computational and in vitro approaches to eliminate animal use from drug discovery for humans and other species. Our immediate objective is to employ in silico ligand protein docking to discover existing drugs (repurposing) and/or new drug candidates capable of inhibiting host proteins involved in infection pathways for the COVID-19 virus, SARS-CoV-2. Our docking targets include four serine hydrolases: TMPRSS2, a protease that activates the SARS-CoV-2 spike protein to enable viral entry into cells; and PLA2G2D, PLA2G4A, and PLA2G6C (NTE/PNPLA6), phospholipases that promote viral replication. We have a crystal structure for PLA2G4A, and we will create homology models for PLA2G2D and PLA2G6C (NTE/PNPLA6). Using these targets, we will reversibly dock approximately 40,000 ligands from the Binding Database comprising FDA-approved drugs along with serine protease and PLA2 inhibitors, including organoboron compounds. Next, we will conduct covalent docking on a ligand subset containing pharmacophores capable of covalently binding serine hydrolases. Consensus ranking from four docking programs will be used to generate a penultimate list of candidate compounds. Those showing high predicted potency against off-target serine hydrolases (acetylcholinesterase, butyrylcholinesterase, carboxylesterase-1, and thrombin) will be excluded. Our final list of compounds will be made publicly available for further evaluation in bioassays.

Department of Bioengineering, Stanford University, Stanford, CA
Animal-free and large-scale generation of microvascularized cardiac organoids for high-throughput bioprinting of human cardiac tissues
The availability of patient-specific cardiac tissue models would reduce our dependency on animal models for preclinical screens and ensure highly predictive human-relevant data for clinical translatability. 3D bioprinting is a uniquely powerful and versatile tool for designing such complex human tissues, yet it is critically limited in its ability to manufacture reproducible arrays of densely-cellular, vascularized, and instrumented cardiac tissue at a scale that is required for high-throughput pharmaceutical screens. Furthermore, owing to the challenges of manufacturing stem-cell derived human tissue at the billion-cell scale, many existing large-scale 3D bioprinting methods rely on primary animal cells and use poorly defined animal-derived media products, and hence fail to avert the suffering of research animals. Here, we propose the development of a xeno-free and ethical bio-ink composed of densely packed microvascularized cardiac organoids. Next, we will apply our latest multimaterial multinozzle printing technology to manufacture instrumented cardiac tissue arrays. Once printed, these organoids will fuse to become densely cellular, aligned, and microvascularized human cardiac tissues that are attached to force sensors for drug screening purposes. The resulting animal-free cardiac tissue biofabrication pipeline is uniquely programmable and scalable, and could greatly accelerate the adoption of ethical, scientifically relevant human-specific cardiac preclinical screens.

Lung Bioengineering and Regeneration, Department of Experimental Medical Sciences, Lund University, Lund, Sweden
Cryopreservation of human precision-cut lung slices for on-demand use in high throughput disease modeling and drug screening assays
Epidemics and pandemics, such as the ongoing COVID-19 outbreak, are predicted to increase due to climate change, globalization and urbanization. Many of the recent epidemics originated from pathogens which did not induce severe disease in animals, underlining major differences between animals and humans. This also highlights the urgent need to develop models of human disease. We have recently developed a 3D model of human lung tissue by generating precision cut lung slices (PCLS) from explanted human lung. PCLS can be grown in the lab for up to one week to model human lung disease and test potential therapies. Despite increased interest in their usage, PCLS have not been widely adopted due to limited access to human tissue at institutions capable of generating PCLS. We aim to develop techniques to cryopreserve PCLS for on-demand testing of pathogens and evaluation of potential therapies. Models of human lung tissue are especially critical to develop as deaths in the majority of recent epidemics were due to acute lung disease. As even accelerated vaccine development takes time, access to human disease models are urgently needed to test potential therapies. Cryopreservation of PCLS has the potential to replace animal studies in this growing area.

Haskins Laboratories, Department of Chemistry and Physical Sciences, Pace University, New York, NY
Toxoplasma gondii: Hollow Fiber Bioreactor Replacement of Mouse Models
Toxoplasma gondii has a complex life cycle with asexual (tachyzoites, bradyzoites) and sexual (sporozoites) stages. Sporozoites are found inside oocysts and expelled in cat feces. This environmentally resistant stage is responsible for disease transmission. T. gondii infects multiple mammalian hosts, estimates suggest that 11 percent of the US population has been exposed. Cats are the only known host where T. gondii completes the sexual cycle. Host specificity is due to the absence of delta-6 desaturase in the cat intestinal epithelium. All culture models for the generation of sexual stages use animals and no robust in vitro model for producing oocysts exists, placing the burden for study of the infective-stage on animal models. Using my experience in developing in vitro culture methods for other intestinal parasites, notably Cryptosporidium parvum, I will develop a completely animal-free culture method employing hollow fiber biotechnology. This methodology will employ the R's of alternatives to animal research: replace existing animal models; refine current culture methods by producing a robust method for parasite culture; and reduce the use of animals in basic science research. area.

This grant is jointly funded by the International Fund for Ethical Research (IFER), Chicago, IL.

The Foundation wishes to thank all applicants who submitted proposals and others for their interest in developing alternative methods of conducting high quality scientific research.

Past Recipients

Click below to view lists of past grant recipients.