Novel sperm imaging technique could improve cattle, human fertility

URBANA, Ill. – University of Illinois researchers have developed a new technique to determine the fertility of sperm samples in cattle.

“This work is a part of a five-year project to develop dairy cattle that are resistant to heat and diseases in tropical areas. We want to donate these cows to developing countries to increase their food production,” said Matthew B. Wheeler, professor in the Department of Animal Sciences at Illinois.

In order to develop these traits in cattle, the researchers need to determine which sperm samples work best for in vitro fertilization. A novel imaging approach, published in the Proceedings of the National Academy of Sciences, moves that effort forward.

“Although males may have sperm that are seemingly perfect, there could be morphological or DNA issues. This approach allows us to evaluate the spermatozoa and select the best in terms of fertility,” said Marcello Rubessa, a research assistant professor in Wheeler’s team.

Traditional techniques for imaging sperm samples are slow and labor intensive, and involve toxic stains. To circumvent this issue, Wheeler’s team, along with a group based in the Beckman Institute for Advanced Science and Technology, used label-free imaging techniques developed in the Beckman Institute’s Quantitative Light Imaging Laboratory(QLIL) to determine what parameters of the sperm make them fertile.

“We knew from the fertilization experiments which sperm samples worked. We used our imaging technique to understand what parameters were important for success,” said Mikhail Kandel, a graduate student with the QLIL. “We saw that the relationship between the size of the head and the tail of the sperm is an important parameter for fertility.”

Additionally, the researchers also improved the speed of the technique. “We used artificial intelligence to automate the process of analyzing these sperm cells,” said Yuchen He, a graduate student with QLIL.

The researchers hope to improve the speed of the technique for future analysis. “The motility of the sperm is sometimes fast. Therefore, we need to do the measurements quickly,” said Gabriel Popescu, director of theQLIL and professor in the departments of electrical and computer engineering and bioengineering at Illinois.

“For many years, we have developed various techniques for label-free imaging knowing that we had to give away molecular specificity,” Popescu said. “However, our newly developed phase imaging with computational specificity brings back the molecular specificity via AI, which is harmless and works on live cells. The applications are limitless, but one that truly benefits from absence of chemical stains is assisted reproduction, as described in this collaborative study.”

The researchers hope to further develop the technique for assisted reproductive technology in humans.

The study, “Reproductive outcomes predicted by phase imaging with computational specificity of spermatozoon ultrastructure,” is published in the Proceedings of the National Academy of Sciences [DOI: 10.1073/pnas.2001754117]. The study was supported by grants from the Ross Foundation, the United States Department of Agriculture, the National Institutes of Health, and the Integrated Grants Management System.

The Department of Animal Sciences is in the College of Agricultural, Consumer and Environmental Sciences at the University of Illinois.

https://aces.illinois.edu/news/novel-sperm-imaging-technique-could-improve-cattle-human-fertility.

217-300-2435

Pigs push forward quick solution for emergency ventilators

ACES News: https://aces.illinois.edu/news/pigs-push-forward-quick-solution-emergency-ventilators

URBANA, Ill. – When Matt Wheeler got the call on a Sunday morning in March – just two days after Gov. J.B. Pritzker issued his first stay-at-home order – he wasn’t expecting to launch an experiment that could save countless lives.

On the call, leaders from the Illinois RapidVent team explained they had built a prototype of an emergency ventilator to address a nationwide shortage amid the COVID-19 pandemic. Laboratory testing looked promising, but the University of Illinois team wanted to understand whether the device worked in animals. Wheeler, who has built and tested lifesaving medical devices in animals, was the obvious choice to join the team.

Within a week, Wheeler wrote a protocol; obtained approval from the Illinois Institutional Animal Care and Use Committee (IACUC); assembled his team, supplies, and animals; and had completed the first 24-hour tests of the ventilator. A few tweaks and a few days later, final testing was complete.

The RapidVent worked.

“If this device saves one person, we did our job. Hopefully it’ll save a whole lot more than that,” says Wheeler, professor in the Department of Animal Sciences at U of I and affiliate in the Department of Bioengineering, Department of Veterinary Clinical Medicine, Carl R. Woese Institute for Genomic Biology, Beckman Institute for Advanced Science and Technology, and the Carle Illinois College of Medicine.

Wheeler’s team tested the device in pigs, widely recognized as the non-primate mammals most physiologically similar to humans.

“Typically the size pig we use for this kind of work is somewhere in the 200- to 250-pound range. The lungs of those pigs are about the same size as a 150-pound human,” Wheeler says.

The team – in head-to-toe personal protective equipment – humanely sedated, intubated, and monitored the pigs as the RapidVent took over the job of breathing. The first test ran for three hours, just to make sure the setup worked. The next step was testing the device on multiple pigs for a full 24-hour period. Using data from these tests, the RapidVent team made a few critical adjustments to the prototype. A few days later, a final four-hour stint rounded out the testing.

The device is designed for short-term, emergency respiratory support in hospitals when regular ventilators are not available. First responders also can hook the device to an oxygen tank to breathe for rural patients during long treks to the nearest hospital.

The product’s need and impact show little sign of slowing down. More than 50 companies have now licensed the design for the Illinois RapidVent and are exploring manufacturing options. When the time comes, Wheeler’s team and his pigs stand ready to test a commercial product.

Wheeler points to the College of Agricultural, Consumer and Environmental Sciences’ (ACES) Imported Swine Research Lab (ISRL) for the experiment’s rapid-turnaround success.

“We could do this so fast because we were already set up with animals and a state-of-the-art biomedical unit managed by the animal sciences department in the College of ACES. Had we not had that facility there, there’s no way we could have done it as quickly as we did,” he says.

Pigs from the ISRL have helped test devices that have saved infants and rebuilt facial bones of injured soldiers, outcomes Wheeler is proud of. But his primary gig is agriculture. Broadly, he and his team work to improve production characteristics in swine and cattle using advanced tools such as gene editing, embryo transfer, and stem cell therapies.

Wheeler’s foundation in agriculture led him to leap with both feet into a project that could save human lives.

“I signed up in ag more than 40 years ago to feed people, to take care of people, and help people who needed help,” Wheeler says. “And so this is just another example of stepping up where we could help; we were ready when the call came in. That’s what we do in agriculture, and what we do in the College of ACES.”

217-300-2435

Wheeler Lab Leads Animal Testing of the Illinois RapidVent Ventilator

The Illinois RapidVent is a working prototype of an emergency ventilator for COVID-19 patients.  Website: https://rapidvent.grainger.illinois.edu/index.asp

Full Press Release: https://rapidvent.grainger.illinois.edu/pr

The United States is expecting a severe shortage of ventilators to help people suffering from the most serious cases of COVID-19. On March 16, 2020, a team of more than 40 engineers, doctors, medical professionals, designers, and manufacturing experts from industry launched an Apollo 13-style project to help address that need.

“Our team is living the Apollo 13 movie,” said William King, the overall project leader. “We have dropped everything else to work around the clock to help respond to the COVID-19 crisis.” King is a Professor of Mechanical Science and Engineering who holds appointments in The Grainger College of Engineering and the Carle Illinois College of Medicine.

“We have a team of brilliant and dedicated people that made something that actually works in less than one week. It’s very inspiring. We hope that we can engage even more people to work on the global response to COVID-19 as we continue to develop the prototype.”

“This Coronavirus can impact a patient’s lungs, and those who are sickest may need help breathing,” said Karen White, MD, PhD, an intensivist at Carle Foundation Hospital and a faculty member in the Carle Illinois College of Medicine. “Ventilators are necessary to help patients get more oxygen. That’s why we’re optimistic that by further developing the Illinois RapidVent we can develop more options for our sickest patients.”

Animal Studies: Animal tests were conducted in order to validate the RapidVent and to evaluate the potential for use in humans infected with COVID-19.The objective of the animal studies was to test and validate the use of a rapid prototyped emergency gas powered ventilator in pigs that is designed after an approved, commercially available ventilator for potential use in humans infected with COVID-19 virus. This animal study represented a critical testing step to build confidence in the design and ultimately support the effort to explore approval for this ventilator for human use. Since the end goal was for these ventilators to be used in human patients, pigs were of special interest because the size of their lungs is comparable to that of humans.

Animal Team Roster: https://rapidvent.grainger.illinois.edu/rosters

Wheeler Lab Team Collaborators:                                                                               Dr. Clifford Shipley, DVM, Dr. Marcello Rubessa, Dr. Derek J. Milner, Dr. Paula Marchioretto, DVM, Mr. Jonathan Mosley, Ms. Sarah Womack, Ms. Sierra Long, and Ms. Jacqueline Newman.

AACUP Collaborators:                                                                                                    Dr. Courtney Hayes and Dr. Nicole Herndon

First 3/4 Gyr Calves Born at Chessie Creek Farm Project

The first 3/4 Gyr calves were born at Chessie Creek Farm in South Carolina in October 2019. The University of Illinois in collaboration with Chessie Creek Farm produced the 3/4 Gyr calves by in vitro fertilization of oocytes collected by our team from 1/2 blood Holstein-Gyr donors. This generation is the next step in the production of a Girolando cattle herd in the United States. This work is supported by a generous gift from the Ross Foundation.

Dr. Wheeler Among Medical College Inaugural 100 Faculty

DR. WHEELER AMONG MEDICAL COLLEGE INAUGURAL 100 FACULTY

Dr. Wheeler, of the Department of Animal Sciences, is among the 100 prominent researchers, administrators, and medical professionals named to the faculty of the new Carle Illinois College of Medicine, the world’s first engineering-based college of medicine.

The medical college is a partnership between the University of Illinois at Urbana-Champaign and Carle Health System, based in Urbana. The college will welcome its first class of 32 students in 2018.

View the full list of inaugural faculty at the medical college: go.illinois.edu/First100MedFaculty.

Holstein X Gyr F1 Hefiers Arrive at Chessie Creek Farm- Animals Will Be Used to Produce the Next Generation of Tropical Adapted Dairy Cattle

Holstein X Gyr F1 Hefiers

Friday, June 15th, 2018 marked the next step in producing more milk for people in developing tropical countries. Fifty-four Holstein X Gyr F1 heifers arrived at Chessie Creek Farm in South Carolina. The animals are part of the Chessie Creek Farm-University of Illinois Project for the Genetic Improvement of Livestock led by Dr. Matt Wheeler and his colleagues. These animals will provide the genetics for the next generation of embryo donors for this project. Early in the fall some of these animals will calve and the dams will provide the first production data that will ultimately enable the selection of superior genetics to disperse to developing countries. The partnership with Chessie Creek Farm has enabled the fast pace of progress toward the goal of disseminating tropical-adapted dairy genetics worldwide. The University of Illinois wishes to thank the owner and all the staff of Chessie Creek Farm for their support and diligent work on this project.

Matthew B. Wheeler

Gradient Light Interference Microscopy (GLIM) judged one of the ten best microscopy innovations in 2018 by Microscopy Today!

GLIM Imaging for Thick Embryos

Gradient Light Interference Microscopy (GLIM) was judged one of the ten best microscopy innovations in the 2018 Microscopy Today Innovation Award competition. Dr. Marcello Rubessa and Dr. Matthew B. Wheeler and their Beckmann Institute collaborators, Dr. Gabriel Popescu, Dr. Tan Nguyen and Ph.D. student Mikhail Kandel will receive this prestigious award at the Microscopy & Microanalysis 2018 meeting in Baltimore, Maryland, on August 5, 2018.

GLIM extracts extract three-dimensional information from both thin and thick unlabeled specimens. GLIM can potentially become a valuable tool for in vitro fertilization, where contrast agents and fluorophores may impact the viability of the embryo. Since GLIM is implemented as an add-on module to an existing inverted microscope, we anticipate that it will be adopted rapidly by the biological community.

Matthew B. Wheeler

First Tropical Adapted Dairy Calves Born at Chessie Creek Farm in South Carolina!

1/2 Blood Holstein X Gyr Female Calves at Chessie Creek Farm

The first 1/2 blood Holstein X Gyr calves were born in South Carolina in March 2018. These calves will ultimately be the donors that will produce oocytes and embryos for donation to feed developing countries. Chessie Creek Farm is the hub of this work which is a collaboration between Chessie Creek Farm and the University of Illinois through the “Genetic Improvement of Livestock Project”. The project aim is to donate large numbers of Tropical-Adapted cattle embryos to the developing world by 2022. The first donation is being negotiated with Tanzania and will likely occur in the Fall of 2018.

Myogenic potential of mesenchymal stem cells isolated from porcine adipose tissue.

Advances in stem cell biology and materials science have provided a basis for developing tissue engineering methods to repair muscle injury. Among stem cell populations with potential to aid muscle repair, adipose-derived mesenchymal stem cells (ASC) hold great promise. To evaluate the possibility of using porcine ASC for muscle regeneration studies, we co-cultured porcine ASC with murine C2C12 myoblasts. These experiments demonstrated that porcine ASC display significant myogenic potential. Co-culture of ASC expressing green fluorescent protein (GFP) with C2C12 cells resulted in GFP+ myotube formation, indicating fusion of ASC with myoblasts to form myotubes. The presence of porcine lamin A/C positive nuclei in myotubes and RTqPCR analysis of porcine myogenin and desmin expression confirmed that myotube nuclei derived from ASC contribute to muscle gene expression. Co-culturing GFP+ASC with porcine satellite cells demonstrated enhanced myogenic capability of ASC, as the percentage of labeled myotubes increased compared to mouse co-cultures. Enhancing myogenic potential of ASC through soluble factor treatment or expansion of ASC with innate myogenic capacity should allow for their therapeutic use to regenerate muscle tissue lost to disease or injury.

Culture in the presence of differentiating porcine satellite cells can initiate myogenesis in porcine ASC prior to fusion with myotubes. Co-cultures of porcine GFP+ ASC with porcine satellite cells undergoing myogenesis exhibit the presence of individual ASC expressing sarcomeric MHC. a Merged fluorescent images for GFP (green), sarcomeric MHC (red) and dapi (blue) clearly show an individual GFP+ mononuclear cell (arrow) expressing sarcomeric MHC, indicating that this is an ASC that has initiated the myogenic program. Also note the presence of a large GFP+ myotube (*) exhibiting clusters of GFP+ nuclei (arrowhead), indicating that this myotube has been formed with contribution of several ASC. b Enlarged single channel images and merged image of the mononuclear ASC marked by the arrow in (a). The arrow in each panel points to the single nucleus of the cell. Note the presence of GFP label in the nucleus. c Enlarged single channel images and merged image showing a cluster of GFP+ nuclei derived from ASC in the region marked by the arrowhead in (a). The arrow in each panel points to a nucleus derived from a porcine satellite cell lacking GFP

Pictured above: Culture in the presence of differentiating porcine satellite cells can initiate myogenesis in porcine ASC prior to fusion with myotubes. Co-cultures of porcine GFP+ ASC with porcine satellite cells undergoing myogenesis exhibit the presence of individual ASC expressing sarcomeric MHC. a Merged fluorescent images for GFP (green), sarcomeric MHC (red) and dapi (blue) clearly show an individual GFP+ mononuclear cell (arrow) expressing sarcomeric MHC, indicating that this is an ASC that has initiated the myogenic program. Also note the presence of a large GFP+ myotube (*) exhibiting clusters of GFP+ nuclei (arrowhead), indicating that this myotube has been formed with contribution of several ASC. b Enlarged single channel images and merged image of the mononuclear ASC marked by the arrow in (a). The arrow in each panel points to the single nucleus of the cell. Note the presence of GFP label in the nucleus. c Enlarged single channel images and merged image showing a cluster of GFP+ nuclei derived from ASC in the region marked by the arrowhead in (a). The arrow in each panel points to a nucleus derived from a porcine satellite cell lacking GFP

Derek J. Milner, Massimo Bionaz & Elisa Monaco, Jo Ann Cameron and Matthew B. Wheeler