BD Biosciences Research Grants

Winter 2014 Research Grant Recipients Talk About Their Research


Abraham Al-Ahmad, PhD
Assistant Professor
Texas Tech University Health Sciences Center, School of Pharmacy

Abstract Title:
Development of a Novel Human Blood-Brain Barrier In Vitro Model as a Drug Screening Platform for Novel Neuroprotective Agents


BD: What is your educational background?

Abraham Al-Ahmad: I hold a BS in biochemistry and an MS in physiology and pharmacology, both from the University of Strasbourg, France. Because we shared half our curriculum with the neurosciences department, I am trained in both neurosciences and pharmacology. I became involved in blood-brain barrier research at the University of Zurich, Switzerland, under the mentorship of Dr. Omolara Ogunshola. There I pursued my interest for cell culture research and developed a novel in vitro model of the blood-brain barrier using two-dimensional and three-dimensional rat cell cultures. After my PhD, I conducted stroke research with Dr. Gregory Bix at Texas A&M Health Science Center, where I demonstrated that domain V, the C-terminal domain of perlecan (a proteoglycan coating the cerebral blood vessels) is released following stroke injury. I accepted a second postdoctoral position with Dr. Eric Shusta at the University of Wisconsin, Madison. Dr. Shusta is a well-respected expert on the blood-brain barrier.

BD: How and when did you become interested in science?

Abraham Al-Ahmad: I was into science very early on, growing up with Star Wars and Japanese science fiction cartoons on French TV. When I was very young, a genetic disease like Duchenne muscular dystrophy was a death sentence. One day at a public swimming pool I met a child in an electric wheel chair. We raced—me swimming and he pool-side in his motorized chair. He won, but that was the moment when I knew by then I will be a scientist who would seek cures for individuals like that boy. I thought of becoming a physician, but after studying biology and pharmacology I decided to pursue science full-time instead.

BD: How did you become interested in your broad field of study?

Abraham Al-Ahmad: I entered stem cell research through a fortunate accident. During my postdoc in stroke research I became interested in cellular events. At the time, Dr. Eric Shusta at Wisconsin was looking for a postdoc to continue development of a human-based blood-brain barrier model developed from pluripotent stem cells. I was experienced with the blood-brain barrier, but I had never worked with stem cells. What fascinated me was the potential to coax these cells to become any cell type, and that these cells—induced pluripotent stem cells—could be harvested from patients.

BD: Describe the project for which you received the BD grant.

Abraham Al-Ahmad: Stroke constitutes the fourth-leading cause of death in industrialized countries and is responsible of one of the highest morbidity rates for survivors. The personal, societal, and economic effects are huge. Despite efforts for discovering novel therapies, there are still no efficient stroke treatments. The only FDA-approved drug, tissue plasminogen activator (tPA), is limited to administration six hours after stroke onset, and the drug carries a high risk of cerebral hemorrhage.

My approach involves using human induced pluripotent stem cells (iPSCs) to model the human neurovascular unit that includes the blood-brain barrier, astrocytes, and neurons. The advantage of iPSCs over embryonic stem cells is that they provide a patient-specific source of cells. During my postdoctoral training with Prof. Shusta, we established a functional in vitro model of the human blood-brain barrier using both iPSCs and embryonic stem cells.

I plan to validate this model to show that it is suitable for screening novel stroke neuroprotective drug candidates. To model stroke injury, we will put our cells into an "oxygen-glucose deprivation" (OGD) stress. This method focuses on depleting cells from oxygen and from glucose supply to mimic similar effect to stroke injury. In addition to the OGD stress, we will also mimic the "reperfusion" injury as it happens once neurosurgeons remove the clot in stroke patients. It is known that the rapid influx of oxygen and glucose is perceived as hyperoxic and hyperglycemic stress. We hope to show that our model responds to OGD stress by cells losing their barrier function. We would also like to investigate how iPSC-derived astrocyte and neuron co-cultures respond to OGD stress, in particular how OGD/reperfusion trigger neuronal cell death.

BD: What are the long- and short-term scientific goals of this project?

Abraham Al-Ahmad: Short-term, we would like to demonstrate proof-of-concept for our model of the neurovascular unit to help discover drug candidates for clinical evaluation. If our system behaves similarly to in vivo models in terms of blood-brain barrier breakdown and neuronal cell death, it will also help us understand the mechanisms of action of novel stroke neuroprotective agents. Long-term, we hope that our model will advance development of a high-throughput screening platform. Instead of spending months to test one small molecule at the time, we may be able to test a dozen molecules per day.

BD: What are the implications of your project for human health?

Abraham Al-Ahmad: I look forward to the day when we can use iPSCs from our own skin cells to better predict the ability to recover from stroke injury and to predict how our individual blood-brain barriers will allow us to cope with neurological diseases. If successful, our model could be a big game changer. Perhaps one day we might have a "blood-brain barrier on a chip" to predict which drugs will cross the blood-brain barrier and also predict the risk of a brain's sensitivity to environmental pollutants

BD: Which BD reagents do you plan to use, and for what purposes?

Abraham Al-Ahmad: We will use mostly flow cytometry or cell imaging reagents, including primary antibodies directed against GFAP (an astrocyte marker), nestin (a neuronal stem cell marker), and beta-III tubulin (a differentiated neuron marker) to validate my ability to differentiate iPSC-derived neural stem cells into neurons and astrocytes. Also primary antibodies targeting blood-brain barrier cells, apoptotic cells, and IgG isotype controls for my primary antibodies.


Enrique A. Armijo
Graduate Student
University of Texas Health Science Center

Abstract Title:
Development of a New Mouse Model Harboring Human Neurons to Study Alzheimer's Disease In Vivo


BD: What is your educational background?

Enrique Armijo: I received my undergraduate degree in molecular biotechnology engineering and my master's degree in neuroscience at the University of Chile, Santiago, where I studied neuronal iron metabolism in Parkinson's disease. In 2011, I moved to the United States to join the laboratory of Dr. Claudio Soto at the University of Texas Health Science Center at Houston, where I currently work. Here I am learning about Alzheimer's disease pathology and its devastating consequences for patients and their families. My current graduate research project involves the use of induced pluripotent stem cells (iPSCs) to model and treat Alzheimer's disease. I expect to receive my PhD in biomedicine in the summer of 2015 from the University of Los Andes, Santiago, Chile, and the University of Texas at Houston.

BD: How and when did you become interested in science?

Enrique Armijo: Throughout high school I was particularly interested in biology and initially planned on a career in dentistry. However, upon my admission to the molecular biotechnology program, I discovered the amazing worlds of cellular and molecular biology and decided to pursue those subjects. That was one of the best decisions in my life.

BD: How did you become interested in your broad field of study?

Enrique Armijo: My interest in stem cells began while I was working on my master's thesis in Chile, when I decided to switch from Parkinson's-related iron metabolism to stem cells. I was curious about the potential use of stem cells for treating humans with neurodegenerative diseases. After learning more about stem cells and iPSCs, I became interested in the generation of iPSCs from patients with Alzheimer's. This technique offers an exciting new opportunity to regenerate patient-specific cells and model the disease in vitro and in vivo.

BD: Describe the project for which you received the BD grant.

Enrique Armijo: Development of new, innovative, and more reliable experimental models is an urgent medical priority. We are developing a new chimeric mouse model harboring human neurons to study Alzheimer's in vivo. This approach offers a unique opportunity to study Alzheimer's pathology through human neurons in the mouse brain, in particular aspects of the human disease that have not been reproduced successfully in previous animal models, including clinical symptoms, mature protein aggregates, and neuronal damage.

This project explores two recently developed techniques in a novel application—iPSCs and the generation of chimeric mice harboring human nerve cells. Until now, nobody has tried to combine these technical developments together to generate a better model of AD.

Our animal models consist of mouse embryos at embryonic stage 14 and neonate mice from wild-type and transgenic mouse models of Alzheimer's. We will inject them with human iPSCs or iPSC-derived neural precursor cells originating from AD patients and fibroblasts from control individuals. Interestingly, neurons generated from iPSCs derived from individuals with Alzheimer's disease show intra- and extracellular aggregates, higher levels of amyloid beta peptide, and hyper-phosphorylated tau protein, recapitulating some key aspects of Alzheimer's in vitro. We plan on reproducing some of the key aspects of Alzheimer's pathology in an in vivo context after grafting human iPSCs in the mouse brain.

BD: What are the long- and short-term scientific goals of this project?

Enrique Armijo: We will work short-term on optimizing our grafting technique to ensure a high survival rate for human cells implanted into mouse brains. We will evaluate human cell survival and differentiation through histological techniques. Long-term, we hope to transplant human cells derived from patients affected by sporadic and familial forms of Alzheimer's into a wild-type mouse to evaluate if the host tissue is affected by the presence of unhealthy human cells. Similarly, we will introduce human cells derived from healthy individuals in the brain of mouse model that develops Alzheimer's-related abnormalities. We will study these abnormalities in the grafted cells and host tissue, with special attention on the abnormalities absent in the animal models—for example, extensive neuronal death and cerebral atrophy.

BD: What are the implications of your project for human health?

Enrique Armijo: Alzheimer's disease is not a normal part of aging, yet approximately 35 million individuals worldwide and more than 5 million Americans suffer from the disease. In societies with increasing life expectancy, such as the United States, the prevalence of Alzheimer's is expected to double over the next 40 years. The disease exacts tremendous socio-political and economic burdens due to a lack of effective therapies. Although numerous in vitro and in vivo models have emerged during past decades, current treatments are palliative and neither slow down nor halt disease progression. This is because current models do not replicate the entire disease, and represent only an incomplete Alzheimer's phenotype.

Developing and characterizing our model of Alzheimer's disease in mice harboring human cells could provide a novel platform for investigating the molecular bases of neurodegenerative processes, discovering new pharmaceutical targets and biomarkers, and developing of new drugs to treat or even prevent the onset of Alzheimer's and potentially for developing new and more relevant models for other brain diseases.

BD: Which BD reagents do you plan to use, and for what purposes?

Enrique Armijo: For in vitro studies, we will use BD antibodies to characterize and isolate our human iPSCs and neural precursor cells. We also expect to order cell culture media, growth factor supplements, plasticware, pipets, and Matrigel® matrix for expanding and differentiating our cells. For in vivo studies, the phenotype of the human grafted cells in the mouse brain will be determined largely by BD antibodies and assay kits. We will study inflammatory/anti-inflammatory responses, apoptosis, and cell proliferation using ELISA assays.


Benjamin Bakondi, PhD
Post-Doctoral Researcher

Abstract Title:
Cell Surface Epitope Expression Analysis of Mobilized Endogenous Bone Marrow Stem Cells for the Treatment of Optic Nerve Crush Injury


BD: What is your educational background?

Benjamin Bakondi: I received my BS in biology from the University of Massachusetts, Amherst. After graduation, I held research positions at Massachusetts General Hospital, where I studied Alzheimer's disease genetics, and at Cornell University Medical College. My work at Cornell involved neural stem cell therapeutics. I spent two years at Columbia University earning my master's degree in biotechnology before entering the University of Vermont's College of Medicine for my PhD. My focus in the lab of Prof. Jeffrey Spees at UVM was to help develop and characterize peptide biologics derived from bone marrow mesenchymal stem cells for the treatment of stroke, heart attack, and peripheral nerve crush injury. I am currently a post-doctoral scientist in the Regenerative Medicine Institute at Cedars-Sinai Medical Center, Los Angeles, where I'm helping to develop stem cell and gene therapy treatments for retinal degenerative diseases.

BD: How and when did you become interested in science?

Benjamin Bakondi: I remember at age eight telling my father that I wanted to be an inventor. I was always taking things apart and trying to figure out how they were made. My father, who was good at chemistry during his college days, was a model for my scientific aspirations. I entered college as a chemistry major, but switched to biology during my junior year. There were times during biology lectures when I believed the class had ended after what seemed like ten minutes when actually for me the time had just flown by. This was a clear sign for me that I should pursue biology research.

BD: How did you become interested in your broad field of study?

Benjamin Bakondi: My interests in biology became focused on research to help those with diseases and disabilities because I felt that the development of treatments is inevitable as long as enough people were working toward that goal. Stem cell research seemed to be the best way to achieve those goals. Although I had a couple of laboratory internships in college, it wasn't until my after graduation and landing a technician position in Dr. Steve Goldman's lab that I had a chance to see how stem cell research is done first-hand. The objective of that work was to help manipulate stem cell behavior and evaluate if the response is of therapeutic use.

BD: Describe the project for which you received the BD grant.

Benjamin Bakondi: All throughout grad school I studied how bone marrow stem cells (BMSCs) provide benefit after acute injuries such as stroke, heart attack, and peripheral nerve crush. Dozens of investigators have used BMSCs to treat such injuries in animal models with success, but to this day nobody's sure how these cells work. It's mostly because the cells don't last long after transplant. BMSCs initially get trapped in the lungs and blood-processing organs after intravenous injection and are mostly undetectable after a couple of weeks. But the benefits last much longer, even for the lifetime of the animal in some cases. How the temporary presence of these cells provides long-lasting benefits is what we will investigate with this grant.

We hypothesized that systemically injected BMSCs incite the mobilization of tissue-protective cells from the bone marrow. We know that endogenous BMSCs and their cell-derivatives regulate multiple aspects of hematopoiesis, among other roles in vivo. So it's not too much of a stretch to suggest that the injected cells could do the same; that some of them reach the bone marrow and influence the activation of endogenous cells that are the ones providing the long-lasting benefits we see.

In our lab, animals that were infused with pre-labeled BMSCs after optic nerve crush had decreased numbers of marrow cells with the p75 surface receptor—the low-affinity nerve growth factor receptor—and the proliferation marker, Ki67, compared with animals that just had the injury. These cells may leave the bone marrow to act beneficially at injury sites. To test this hypothesis, we will characterize the mobilization kinetics of these cells as they travel from the marrow, into the circulation, and arrive at the retina. If the mobilized cells are indeed recruited to the optic nerve, we will characterize their expression of protective factors in situ to determine how they might be influencing the local injury environment as well as the duration of their action over several time points. We will use antibodies for flow cytometry to characterize the surface epitope expression profile of these cells, and can isolate them from the circulation for transplant experiments by fluorescence activated cell sorting. Later on, if the transplantation of these isolated cells without prior BMSC infusion reduces optic nerve damage, and if blocking the migration of these cells before transplantation shows that we removed this protection, then this would strongly support our hypothesis of how transient BMSCs provide long-term benefits.

BD: What are the long- and short-term scientific goals of this project?

Benjamin Bakondi: We first need to identify which population(s) of endogenous cells enters the circulation in response to BMSC injection. This is achieved by defining the unique cell surface epitope expression profile on the mobilized cell populations. The ultimate goal of this study is to identify cell-protection mechanisms and demonstrate the translational potential. The future of such cell transplants in clinical settings may be in the form of off-the-shelf allogeneic cells that are defined by a specific combination of surface epitopes. The better defined these cells are, the more homogeneous the cell population we would have—and this translates to more reproducible and thus predictable outcomes, which means safer therapies. It is also quite possible that different endogenous cell populations are mobilized by different types of injuries, in which case multiple transplant-ready cell populations differing by their epitope expression profile may be uniquely suited to treat certain injuries over others.

BD: What are the implications of your project for human health?

Benjamin Bakondi: Traumatic optic neuropathy results from sudden damage to the optic nerve, often from military combat or automobile accidents, and frequently leads to vision loss. Cell treatments may reduce the severity and extent of vision loss from such injuries in patients. This work has implications for other types of nerve injury occurring through blunt trauma or as a side effect of surgery. Another study showed that BMSC injection improved cavernous nerve survival and function in an animal model for the type of nerve damage that often accompanies prostatectomy in patients. Evidence exists for treating sciatic nerve damage in this manner as well.

BD: Which BD reagents do you plan to use, and for what purposes?

Benjamin Bakondi: We plan to utilize the large selection of BD Pharmingen™ antibodies targeting rat to make immunophenotyping panels for multicolor flow cytometry based on candidate peptides on mobilized cells. Immunohistochemistry will be performed on retinal sections to identify surface antigens, and intracellular signaling and secreted peptides in the mobilized cells. Accompanying reagents and controls for these studies include 7-AAD, binding buffer, antibody diluent, fix buffer, and isotype control antibodies such as IgG2a, IgG2b, and IgG1. Our research may also involve investigating differences in circulating cytokine levels after BMSC transplant, which we can quantify by ELISA using BD OptEIA™ sets.


Abhik Bandyopadhyay, PhD
Research Assistant Professor
University of Texas Health Science Center, San Antonio

Abstract Title:
Identification and Characterization of Human Mammary Stem Cells Susceptible to Initiate Mammary Tumorigenesis During Aging


BD: What is your educational background?

Abhik Bandyopadhyay: I received my BS in chemistry and MS in biochemistry from Calcutta University, India. While working as a biochemist at Ramakrishna Mission Seva Pratishthan Hospital in Calcutta I received my PhD in clinical biochemistry. The hospital is affiliated with the Vivekananda Institute of Medical Sciences and Calcutta University. I moved to the United States in 1987 for my post-doctoral fellowship in the Tumor Biology Laboratory at the University of Nebraska, Lincoln, where I studied mammary gland and cancer biology. I received additional post-doctoral training in molecular drug metabolism and transport at the Pharmacology and Experimental Therapeutics Department, which is part of the College of Pharmacy at the University of Kentucky, Lexington. My appointment as Assistant Professor/Research at the University of Texas Health Science Center, San Antonio, occurred in 2000. My work involves therapeutic targeting of tumor-promoting transforming growth factor beta (TGFβ) in murine cancer models. Since 2006 I have researched the generation of cancer stem cells, mammary stem cell aging, and the development of breast cancer.

BD: How and when did you become interested in science?

Abhik Bandyopadhyay: The mysteries of nature fascinated me from my childhood. Later, in school, I was encouraged to pursue biological sciences by a teacher who noticed my enthusiasm in science projects. Since then, it is a wonderful journey enjoying every day in the exploration of basic and translational research.

BD: How did you become interested in your broad field of study?

Abhik Bandyopadhyay: While studying the efficacy of molecularly targeted therapies in preclinical animal models of breast cancer in the presence or absence of chemotherapy, I noticed that anti-cancer drugs were ineffective in shrinking the tumors after a certain treatment period. At that point, tumors became more aggressive, spreading to different organs and killing the animals. At that time, a new theory emerged via experimental data, namely that a small number of stem-like tumor cells, also called cancer stem cells, escaped therapy and were responsible for creating drug resistance and cancer recurrence. Inhibiting several genes in the stem cell self-renewal pathway in an animal model of late-stage breast cancer resulted in modest success in treating these animals.

I then became interested in learning whether these breast cancer stem cells originated from normal breast stem cells, and how their generation could be prevented. Since age is the number one risk factor for breast cancer, the aging model attracted me in this investigation. During aging, normal stem cells responsible for lifelong tissue maintenance and repair are susceptible to changes in their niche and genomic integrity, which may lead to their possible tumorigenic transformation. Our studies using an animal model, where old mammary stem cells were transplanted to regenerate mammary glands in mice, indicated that these cells were prone to produce pre-cancerous lesions and might be the origin of age-associated breast cancer. These results in mice led me to explore cellular and molecular mechanisms of mammary stem cell dysfunction in progressively aged individuals, and whether these cells transformed themselves to cancer stem cells and subsequently caused breast cancers.

BD: Describe the project for which you received the BD grant.

Abhik Bandyopadhyay: The number of cancer cases, particularly in older people, is expected to increase dramatically over the next two decades. About 80% of all breast cancers arise in women over age 50, with a lifetime risk of 1 in 8. Although mammary stem cell aging is implicated as a possible cause, the efficient isolation and molecular characterization of susceptible dysfunctional human mammary stem cells formed as a result of aging is still lacking. In this study, we will first isolate functional young and old human mammary stem cells through flow cytometry. We expect to establish a unique gene signature for the tumorigenic transformation phenotype of old mammary stem cells using next-generation single-cell whole transcriptome sequencing, and compare these to young mammary stem cells. This information will help us define a possible pharmacological or dietary intervention to prevent the initiation and/or early progression of age-related breast cancer. Next, we will compare young and old mammary stem cell function in terms of their in vivo regeneration ability, and also their potential for pre- and early tumorigenic transformation in immune-deficient mice. We expect to see lesions similar to pre-neoplastic atypical ductal hyperplasia or early neoplastic ductal carcinoma in situ in the regenerated human breast-like bi-layered mammary epithelial structures originated from old mammary stem cells. We have observed this earlier in a mouse model of mammary gland regeneration.

BD: What are the long- and short-term scientific goals of this project?

Abhik Bandyopadhyay: Our short-term goal is to define a unique gene expression signature related to mammary stem cell dysfunction and susceptibility to tumorigenic transformation. We will also compare the unique functional property of young and old human mammary stem cells to regenerate human mammary gland-like structures, and determine their potential to neoplastic transformation. Our long-term goal is to shed light on our understanding of the cellular and molecular pathways that mediate cancerous transformation of aging human mammary stem cells.

BD: What are the implications of your project for human health?

Abhik Bandyopadhyay: This project will set the foundation to delineate clinically relevant novel biomarkers for the prevention, early detection, and treatment of age-associated breast cancer, and thus will promote longevity and healthy aging in women.

BD: Which BD reagents do you plan to use, and for what purposes?

Abhik Bandyopadhyay: We will use BD antibodies for mammary stem cell sorting by flow cytometry, and BD collagen and Matrigel® matrix for in vivo human mammary gland-like structure regeneration in an animal model. We will also order BD reagents for staining, fixing, and preparing cells for flow or immunofluorescence analysis.


Takanori Takebe, MD
Visiting Associate Professor

Abstract Title:
De Novo Transplantable Liver Bud Generation from Human iPSCs


BD: What is your educational background?

Takanori Takebe: After working as a researcher at the Scripps Research Institute Department of Chemistry in 2009, and as an intern at the Columbia University Department of Transplantation Surgery in 2010, I received my medical degree from the Yokohama City University School of Medicine in 2011. The same year I became a Research Associate at the Department of Regenerative Medicine, Yokohama City University. Since my medical school days I have published several papers in leading medical journals on organ bud generation from induced pluripotent stem cells (iPSCs). In these articles, I proposed stem cell–derived organ bud transplantation therapy as a new approach to combat intractable organ failure. My recent appointment as Associate Professor at age 26 makes me one of the youngest faculty members ever in Japan.

I concurrently hold a joint appointment as a visiting associate professor at Stanford University where I'm advancing previous work on organ bud development. I am also affiliated with Mirai Design Lab, sponsored by Dentsu, to establish a new field, Advertising Medicine, for studying the role of communication in next-generation healthcare systems.

BD: How and when did you become interested in science?

Takanori Takebe: I began working in a lab in 2006, during medical school, and never left. As I'm deeply involved in stem cell biology, I was always skeptical about the current exaggerated potential of regenerative medicine. I am now trying to establish a truly effective approach to regenerative medicine, but with improved scientific understanding.

BD: How did you become interested in your broad field of study?

Takanori Takebe: Stem cells and regenerative medicine sounded promising to me from early on. After my time as a surgical intern at Columbia University, a leading liver transplant center, I changed the path of my medical career from clinical surgery to basic scientist. As a transplant physician you're constantly facing patients who are sadly just waiting to die. I felt I could accomplish more by developing an alternative approach to transplantation. To tackle this challenge, I began studying stem cells and regenerative medicine for the treatment of severe organ failure—conditions where effective surgical treatments are currently lacking.

Current protocols for cell differentiation or reprogramming are quite inefficient. They fail to produce therapeutically effective cell populations from iPS or embryonic stem cells. To improve the efficiency and establish effective therapies, we need to understand better the natural differentiation process and adapt that knowledge into stem cell culture. If we think of organogenesis or organ development, the spatiotemporal or four-dimensional multi-cellular interactions are crucial for guiding cellular differentiation. Thus, the questions that currently interest me would be the role of multi-cellular communications and environmental stimuli for targeting cell differentiation from stem cells.

BD: Describe the project for which you received the BD grant.

Takanori Takebe: With co-workers, I recently developed a preclinical model for treating liver failure using human iPSC-derived organ bud transplantation. This work was published in Nature in 2013. Using this technology, a transplantable iPSC-derived organ bud could be self-organized with somatic tissue–derived human stromal progenitors by mimicking the early developmental process. The next critical steps would be to translate this regenerative method into entirely human iPSC-derivatives including stromal progenitors, and to determine their therapeutic potential following transplantation. Here, we hope first to establish methods for generating phenotypically defined stromal progenitors from human iPSC. Second, we plan to optimize the culturing methods of the 3D liver bud through application of differentiated mixed progenitors from human iPSCs. We hope to develop transplantation procedures that maximize the post-transplant functionalization of iPSC-derived liver buds in an immunodeficient animal model.

BD: What are the long- and short-term scientific goals of this project?

Takanori Takebe: Short-term, we expect to establish a method for generating liver bud–forming endothelial and mesenchymal progenitor cells from human iPSCs through modified published protocols. This will involve the use of antibodies and FACS antibodies for phenotypic validation. Long-term, we hope to develop a functional and vascularized human liver from in vitro–grown liver bud generated entirely from human iPSCs.

BD: What are the implications of your project for human health?

Takanori Takebe: More than 100,000 patients die in the United States ever year while waiting for an organ transplant. Human iPSC–derived organ bud technology is the only feasible paradigm for establishing a system for the de novo generation of vascularized, functional organs from iPSCs. Given the unsatisfactory clinical outcomes of the cell-based therapies that are currently the main goal of stem cell therapy, this proof-of-principle, ie, organ bud transplantation therapy, could revolutionize the application of regenerative medicine to the treatment of end-stage organ failure. This work should also provide a unique opportunity to study human developmental biology and disease modeling as well as supply a drug-screening platform.

BD: Which BD reagents do you plan to use, and for what purposes?

Takanori Takebe: We will use mainly antibodies to human iPSC-derived cells. Other reagents and equipment include cell culture matrix and cytokine assays for deriving or confirming targeted stromal progenitor differentiation. We will probably also need BD Biosciences reagents for high-resolution or intravital live imaging, and for detecting gene and protein expression for evaluating liver bud maturation and functionality.



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