Bedford Research Foundation 2011 Newsletter

Read about all of the progress and the research that has occurred at the Foundation over the course of the past year! Dr. Kiessling outlines her vision for the upcoming year as well. Thank you for your support.


Progress in Circadian Rhythms And Stem Cells

BSCRF’s new mouse embryonic stem cells, PL034 (see below for information on the first incubator videomicroscope).

BSCRF scientists have derived two unique lines of stem cells that may lead to a breakthrough in the efficiency of stem cell derivation and expansion.

BSCRF scientists are following up their discovery that the genes that regulate the rhythms of daily life, circadian rhythm genes, may play important roles in stem cell derivation and stability in culture. Circadian rhythm genes regulate cells in the body by turning “on” and “off” over a 24-hour cycle in response to signals such as light/dark cycles, hormone pulses, and body temperature variations.

Currently, stem cells are cultured in constant temperature in the dark. If BSCRF’s research
proves that circadian rhythm genes play important roles in stem cell division and stability, it could markedly improve the efficiency of stem cell derivation and expansion, urgently needed to produce major advances in stem cell therapy.

Circadian Rhythm Genes: turn “on” and “off” in response to the rhythm of daily life.

To conduct this research, foundation scientists are using a genetic technology that links the circadian genes of a mouse with a gene from a firefly. When the circadian gene is “on”, the mouse cells glow like a firefly; when the circadian gene is “off”, the cells go dark. This mouse, “PER2Luc,” was derived by a circadian gene scientist several years ago and has been used by Dr. Fred Davis of Northeastern University, to study circadian gene expression in mouse tissues.

Body temperature of the mouse dips approximately 1 celsius at the beginning and the end of daylight.

BSCRF scientists have derived two new lines of embryonic stem cells from PER2Luc embryos. Light emitted by the stem cells is detectable in Dr. Davis’s luminometer, but BSCRF scientists are developing microscope equipment to record light emitted by individual cells in order to compare standard stem cell culture conditions with new culture conditions that support circadian rhythm such as the temperature variations of a mouse. These new stem cells will also be useful to all scientists seeking to understand the relationship between circadian rhythm and cell functions.

If supporting circadian rhythm genes is important to stem cells, either for their expansion as pluripotent cells or for their uniform differentiation, new methods of stem cell culture need to be developed.

To begin to determine the importance of circadian genes, BSCRF scientists have brought together several different pieces of equipment in order to continually visualize the new PER2Luc stem cells and their emission of light. Because the bioluminescence signal is very low, special cameras are being tested to find one that can detect light emission in individual cells in culture.

The programmable microscope stage incubator (OkoLabs) is being quality controlled by monitoring the cleavage and development of fertilized mouse eggs continuously for six days because mouse embryos require extremely fastidious culture conditions for successful early development. This work has provided new benchmarks for early mouse embryo development in culture to the stages needed for successful stem cell derivation.

The PER2Luc mouse has circadian genes coupled with firefly “Luciferase” genes. When the circadian genes turn “on” they glow, dimly, like a firefly under the microscope.

The 2011 Activated Egg Symposium

2010 Keynote: Janet Rossant,  PhD

Nov 4, 2011: The keynote for our ninth annual symposium is Dr. Janet Rossant, Professor of Molecular Genetics, Obstetrics and Gynecology, University of Toronto, and Chief of Research at the Hospital for Sick Children.

Dr. Rossant is internationally recognized for her pioneering research on stem cells and how they develop into tissues. Dr Rossant will speak on “Manipulating the mouse embryo: from lineages to stem cells and back again.” (see full speaker list at the Symposium Website) The dinner speaker is Chris Hempel, Founder, Addi and Cassi Fund, with compelling talk on the “Regulatory hurdles to research”.

Sponsored by Hamilton Thorne, The McKnight Foundation, Irv & Diane Naylor and Qualcomm.

Testis Stem Cell Project

Thanks to private donations, BSCRF scientists have launched the testis stem cell project.

Phase 1 is the isolation of a new line of testis stem cells from the Per2Luc mouse to study the role of circadian genes in testis stem cells. Phase 2 is to improve the efficiency of deriving testis stem cells from cryopreserved (frozen to stay alive) Per2Luc testis tissues. Phases 1 and 2 are underway.

Phase 3, starting in early 2012, will be to collaborate with Dr. Martin Dym, Georgetown University, in deriving human testis stem cells from cryopreserved biopsies archived in his laboratory. We will compare the efficiency of testis stem cell derivation using our newly developed circadian culture conditions with the efficiency previously reported by Dr. Dym.

Phase 4 will derive patient-specific stem cells from the male volunteers for our study.

CWU Alumni Award

In October, Dr. Kiessling was honored by Central Washington University with the Distinguished Alumni Award. Dr. Kiessling earned her second Bachelor’s and a Master’s degree there. Robert Lowery of CWU praised Dr. Kiessling, “Those degrees, along with a subsequent doctorate in Biochemistry/ Biophysics from Oregon State, became the educational foundation of her extraordinary career…”

UVA Alumni Award

In March, Dr. Kiessling was awarded the first ever Alumni Achievement Award by the University of Virginia School of Nursing. An enthusiastic crowd attended her talk, “Retroviruses, Reproduction, and Regenerative Medicine: The Influence of Federal Funding.” Watch the UVU Alumni Award video.

Victoria Staebler Joins The Foundation Board

The Foundation welcomes Victoria Staebler, Senior Financial Advisor at Merrill Lynch, to the Bedford Foundation Board of Trustees. Vicki holds degrees from Middlebury college and the University of Vermont, serves on the board of directors of The Boston Club, and has honorary board positions at PCI Media Impact, Inc. and Planned Parenthood of Northern New England.

New Lab Staff At The Foundation

We welcome two new staff members: Jacob Noble and Jack Vernamonti. Jake has a BS in physics and math from Northeastern University and is heading up the time-lapse, circadian rhythm, video projects. Jack has a BS in Biochemistry from Colby College and is Coordinator for the Special Program of Assisted Reproduction (SPAR), proudly announcing the birth of 157 babies free of HIV infection.

A Letter From The Director About Research At The Foundation

Our new century has brought world-wide turmoil to social structures, weather patterns, and the earth’s crust itself. Amidst financial collapses, government renewals, tsunamis, hurricanes and earthquakes, people all over the world are redefining “normal.”

So too with biomedical research. Gone is the paradigm of dependence on the federal government for basic research funding: the National Institutes of Health (NIH) can only fund about 10% of submitted grant applications from outside institutions, and that will not change for the foreseeable future. In addition, since funding “sure bets” accounts for most of the federal research budget, speculative research will not be federally funded. And speculative research is where the big breakthroughs lie. As stated by the Pew Charitable Trusts: “If 90% of our funded projects succeed, we are not taking enough risks.”

As the world’s people and institutions struggle with how to do more with less, so must science. American scientists are fortunate in that since the inception of the U.S. tax code in 1917, exemptions have been granted for charitable giving, supporting the notion that citizens have the right to support public causes they believe in.

Despite the global turmoil, our new century has brought steady growth to the Bedford Research Foundation. Our goal is to dramatically improve the efficiency of deriving patient-specific stem cells from naturally pluripotent sources: unfertilized eggs and testis. The added value in doing this is the information we are gaining about embryo development, because to regenerate tissues (heart, spinal cord, immune system, pancreas, etc) we need to study how those tissues form in the mouse. Combined, the work will not only benefit the new field of regenerative medicine, but will also provide new insights into normal human development, perhaps providing avenues to side-step degenerative diseases.

My frustration about how much more we could accomplish each day with more people, more money, is balanced by pride in our ability to do more with less, and by gratitude to the supporters who believe in our mission and our goals, and to the exceptional committee members who meet to support our needs for research oversight. Our administrative costs are low, our ratio of new information/research dollar spent is high, our goals are lofty

With gratitude for your support,

Ann A Kiessling, PhD
Director, Bedford Research Foundation

Who is Bedford Research Foundation?

Philanthropy Is The Key To Continued Progress

The average cost of each experiment is $90,000. Because much of our overhead is covered by fee-for-service laboratory tests, 92% of every dollar donated goes directly toward these experiments. This innovative funding model allows Bedford Research scientists greater flexibility to move quickly in promising new research directions.

Continued progress requires meeting our annual funding goal of $450,000 in 2019.

Donate Today!

You Say Embryo, I Say Parthenote

Julia Galef, Scientific American

 

 

Scientific American November 2011

You Say Embryo, I Say Parthenote“, BSCRF and the importance of parthenote stem cells reported in the November, 2011 Scientific American by Julia Galef.
embryos and parthenotes Scientific America
Parthenogenetic stem cells Image: 
From “Derivation of High-Purity Definitive Endoderm from Human Parthenogenetic Stem Cells using an in vitro Analog of the Primitive Streak,” by Nikolay Turovets et al., in Cell Transplantation, Vol. 20, No. 6; June 2011

Check out the Scientific American article, then for more information, see the BSCRF article, “The History of the Dickey-Wicker Amendment” on the ban on federal funding of non-embryonic, parthenote stem cell research.

And also, see the BSCRF one-minute video, What are parthenote stem cellsWhat Are Parthenote Stem Cells?

Thanks to your donations, Bedford Stem Cell Research Foundation is one of the few laboratories able to work on parthenote, non-embryonic, stem cell research.

Donate Now

 

 

Dr. Janet Rossant to be keynote at 2011 Activated Egg Symposium

We’re delighted to report that Dr. Janet Rossant, Professor of Molecular Genetics, Obstetrics and Gynaecology, University of Toronto, and Chief of Research at the Hospital for Sick Children has graciously agreed to be our keynote speaker for the 2011 Activated Egg Symposium to be held Nov 4, 2011, at the Henderson House in Weston, MA.

Dr. Rossant is internationally recognized for her pioneering research in mouse genetics. Her major findings are related to the question of how genetically identical cells adopt distinct characteristics during embryo development.

In 2010, she received the Premeir’s Summit Award, and they made this video:

A little about more about Dr. Rossant:

Janet Rossant grew up in the UK and trained at the Universities of Oxford and Cambridge. When still a graduate student, she conducted now-classic work defining cell lineages and cell fates in the early mouse embryo. In 1977 Dr. Rossant moved to Canada and joined the faculty at Brock University. From 1985 to 2005, she was a researcher at the Samuel Lunenfeld Research Institute at Mount Sinai Hospital in Toronto. She joined the Hospital for Sick Children in 2005 and became the first female Head of its Research Institute since its founding in 1954. She is also a University Professor in the Department of Molecular Genetics at the University of Toronto.

Throughout her career, she has been a pioneer in manipulating the mouse embryo, deriving novel stem cell lines and interrogating the mouse genome. Most recently, building on her ongoing studies of the mouse blastocyst and the stem cells that arise from it, she is applying her developmental biology skills to derive definitive endoderm lineages from human embryonic stem cells and induced pluripotent stem cells.

As Chair of the Canadian Institutes of Health Research working group on stem cell research and as Deputy Scientific Director of the Canadian Stem Cell Network, Dr. Rossant continues to play a leadership role in setting Canada’s public policy regarding stem cell research.

She has received many accolades for her research, including being elected Fellow of the Royal Society of London (2000), and a Foreign Associate of the National Academies of Science, USA (2008). She was awarded the McLaughlin Medal of the Royal Society of Canada (1998), Eli Lilly/Robert L. Noble Prize from the National Cancer Institute of Canada (2000), Killam Prize for Health Sciences (2004), and FASEB Excellence in Science Award (2004). She received the 2007 March of Dimes Prize in Developmental Biology along with the late Dr Anne McLaren, and the 2007 Conklin Medal of the Society for Developmental Biology, of which she is a Past President.

Here’s another video biography by the Toronto Region Research Alliance in April, 2009:

Bedford presents Post Vasectomy Semen Analysis test kit at Mass Innovation Nights

Foundation Staff | May 24, 2011

 

 

As you may know, Bedford’s laboratory helps cover some of our overhead by offering a couple unique products — GEM and PVSA — that were developed as by-products of our research. PVSA, the newer of the two, was launched just 18 months ago.

PVSA is the first and only post-vasectomy test kit that provides CLIA certified laboratory results from a mail-in kit. This kit was developed using Bedford’s patented specimen fixative and mail-in kits for research.

The kit solves a vital problem for urologists: they are liable for their vasectomy surgeries until their patient’s surgery has been confirmed with two semen specimens.

The kit is inexpensive, easy to use, and helps doctors solve an almost 79% rate of non-compliance from patients. This solution has proven so powerful, we’ve more than tripled our number of clients in the last 10 months.

We are very proud to be presenting this innovative solution at Mass Innovation Nights on June 8, from 6-8:30 PM in Westborough. This event is free and open to the public. Please join us! Also, Mass Innovation Nights will select 4 of the 11 featured products to give a five minute presentation.

 

 

Video: Dr. Ann A. Kiessling receives first Alumni Achievement Award

From the University of Virginia:

The Alumni Achievement Award is a new award established in 2010.  It honors an alumna or alumnus who has shown superior achievement in a field other than nursing since graduating from the University of Virginia School of Nursing.  It recognizes distinction in one’s field and/or contributions to another profession, business or industry, government or public service, education, science or technology, or service to humanity.

2010 ACHIEVEMENT AWARD:  Ann Anderson Kiessling (BSN ’64)

Ann Anderson Kiessling received her BSN from U.Va. in 1964, BS in Chemistry and MS in Organic Chemistry from Central Washington University, and her PhD in Biochemistry from Oregon State University.  During the 1970s, her postdoctoral research explored relationships between viruses and cancer and led to the controversial discovery of Reverse Transcriptase in normal human cells. Believing that reverse transcription may play a role in early embryo development, she began to study eggs, sperm and early cleaving embryos.  Her expertise in reverse transcriptase and the reproductive tract led to the first studies of semen transmission of the AIDS virus in 1983, and in 1985 Harvard Medical School recruited her as Associate Professor in Obstetrics, Gynecology, & Reproductive Biology.  She is currently Associate Professor of Surgery.

In the 1990s, Ann combined her background in human egg biology and AIDS with her interest in stem cell research and founded the Bedford Stem Cell Research Foundation, in response to a controversial clinical research need that was best undertaken by an independent public charity.  The Foundation still receives no federal funding. There Ann developed the first program of assisted reproduction for couples living with HIV disease, and the first human egg donor program for stem cell research, a program that has become a model worldwide.  In 2003, she published Human Embryonic Stem Cells: An Introduction to the Science and Therapeutic Potential, the first textbook on the controversial topic. She has also published more than 100 scientific papers and given more than 60 lectures to audiences around the world. Her writings can be found in publications such as Nature, Lancet, Proceedings of the National Academy of Science and Connecticut Law Review, and she has been the focus of articles in The Boston Globe, Newsweek, and The Wall Street Journal.

Under Ann’s leadership, the Bedford Stem Cell Research Foundation has developed promising advances in deriving patient-specific stem cells while also tackling the moral and ethical issues in stem cell use.  Its programs have helped more than 120 couples affected by HIV disease have safe, healthy babies – and one day may lead to cures for spinal cord injuries, Parkinson’s, and HIV disease.

The First Five Days of a Mouse in GEM, Protein-Free Culture Medium

Foundation scientists are deriving new stem cell lines from fertilized mouse eggs in fully defined, protein-free culture medium, GEM (Gamete Embryo Medium). GEM is a specialized fluid that simulates fluids in the womb. The goal of these experiments is to improve the efficiency of deriving new stem cell lines from testis biopsies. By studying time lapse videos of developing mouse embryos, Foundation scientists will pinpoint the exact time to add stem cell growth factors to the culture medium.

Like all mammalian eggs, fertilized mouse eggs are huge cells that undergo several cleavage divisions before entering the uterus for further development. Each day is marked in this video, watch for these stages:

Day 1: Eggs are fertilized and cleave to the 2-cell stage
Day 2: 4-cell stage
Day 3: Morula (8 to 16 cells)
Day 4: Early blastocysts (40-60 cells)
Day 5: Hatching blastocysts (120-250 cells) This when the embryos could implant in the uterus for further development.

GEM, Bedford Research Foundation’s protein-free, chemically-defined embryo culture medium, supports the development of fertilized mouse eggs to blastocysts, the stage at which embryonic stem cells are derived. This unique culture medium has been more than 20-years in the making and is formulated based on the results of experiments designed to determine the most supportive in vitro conditions for assisted reproduction and embryo manipulation (references). The Foundation has made this medium available for research, academic and educational purposes – more info here.

 

Q & A with the Director about Stem Cell Research at the Foundation

Foundation Director, Ann A. Kiessling, PhD

This article is an excerpt from the Winter Newsletter (pdf).

Q: Has Obama’s Policy Helped Fund Stem Cell Research? 
A: President Obama’s executive order to rescind the restrictions on the number of stem cell lines that could be studied with federal funds was valuable for some studies, but as long as the Dickey-Wicker amendment controls federal funds, the development of stem cells from unfertilized eggs, a prime goal of Bedford Research scientists, cannot be federally funded.

Q: Why do patients need their own stem cells? 
A: The clinical trials that have provided “proof-of-principle” for cell-based therapies, e.g. transplantation of pancreatic cells for diabetes, have revealed that although some cells function normally, many fail because the transferred cels are rejected as “foreign.” The same is true for transplanted bone marrow in cancer therapies. If the stem cells were the patient’s own (i.e. patient-specific), they would not be rejected.

Q: Why do Bedford scientists work with parthenote and testis stem cells? 
A: Parthenote stem cells are derived from unfertilized human eggs, not embryos. Parthenote stem cells behave like embryonic stem cells in the laboratory, multiplying to the trillions needed for therapy, either for the egg donor herself, or for tissue-matched patients. Testis stem cells offer the possibility of deriving stem cells for every man in need, if they can be encouraged to multiply to the trillions possible with parthenote stem cells.

Q: How does Bedford Stem Cell Research Foundation fund its research? 
A: By private donations. The Foundation’s licensed clinical laboratory conducts highly specialized fee-for-service tests which cover the costs of the laboratory infrastructure. This allows research activities to proceed with minimal overhead. The lack of dependence on federal dollars also frees the Foundation from the administrative costs of separate accounting practices required by the Dickey-Wicker amendment.

Evidence for the cure of HIV infection by CCR5 – 32 stem cell transplantation

Kristina Allers,*, Gero Hütter, Jörg Hofmann, Christoph Loddenkemper, Kathrin Rieger, Eckhard Thiel and Thomas Schneider

Dec 2: Timothy Ray Brown can personally affirm that stem cells cure HIV disease, according to a December 2 report in the journal BLOOD. Mr. Brown, an HIV-positive American living in Germany, had leukemia and underwent chemotherapy and bone marrow transplantation in Berlin in 2007. His bone marrow match carried a rare gene mutation in the CCR5 receptor protein, rendering the transplanted cells resistant to HIV infection. Twenty months following the bone marrow transplant, the German team reported Mr. Brown’s leukemia appeared cured, and there was no evidence of HIV in his blood even though he had stopped his antiviral medication prior to the bone marrow transplant.

Now, a year later, the German team has re-examined Mr. Brown’s immune system for evidence of latent HIV infected cells, and found none. The uninfected transplanted bone marrow cells have replaced immune cells in all the parts of Mr. Brown’s body examined. This evidence is consistent with a cure of his HIV disease. Blood, Journal of the American Society of Hematology

This promising proof-of-principle success provides additional support for the patient-specific stem cell based therapy described in Bedford’s June Science Highlights.

ARTICLE ABSTRACT: HIV entry into CD4+ cells requires interaction with a cellular receptor, generally either CCR5 or CXCR4. We have previously reported the case of an HIV-infected patient in whom viral replication remained absent despite discontinuation of antiretroviral therapy after transplantation with CCR5{Delta}32/{Delta}32 stem cells. However, it was expected that the long-lived viral reservoir would lead to HIV rebound and disease progression during the process of immune reconstitution. In the present study, we demonstrate successful reconstitution of CD4+ T cells at the systemic level as well as in the gut mucosal immune system following CCR5{Delta}32/{Delta}32 stem cell transplantation, while the patient remains without any sign of HIV infection. This was observed although recovered CD4+ T cells contain a high proportion of activated memory CD4+ T cells, i.e. the preferential targets of HIV, and are susceptible to productive infection with CXCR4-tropic HIV. Furthermore, during the process of immune reconstitution, we found evidence for the replacement of long-lived host tissue cells with donor-derived cells indicating that the size of the viral reservoir has been reduced over time. In conclusion, our results strongly suggest that cure of HIV has been achieved in this patient.