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GENTAUR Ltd.
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Whetstone London N20 9BH
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San Jose, CA 95123
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6017 Snell Ave, Suite 357
San Jose, CA. 96123
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Antibody Transforms Stem Cells Directly Into Brain Cells
Current techniques for turning patients' marrow cells into cells of some other desired type are relatively cumbersome, risky and effectively confined to the lab dish. The new finding points to the possibility of simpler and safer techniques. Cell therapies derived from patients' own cells are widely expected to be useful in treating spinal cord injuries, strokes and other conditions throughout the body, with little or no risk of immune rejection.
"These results highlight the potential of antibodies as versatile manipulators of cellular functions," said Richard A. Lerner, the Lita Annenberg Hazen Professor of Immunochemistry and institute professor in the Department of Cell and Molecular Biology at TSRI, and principal investigator for the new study. "This is a far cry from the way antibodies used to be thought of -- as molecules that were selected simply for binding and not function."
The researchers discovered the method, reported in the online Early Edition of the Proceedings of the National Academy of Sciences the week of April 22, 2013, while looking for lab-grown antibodies that can activate a growth-stimulating receptor on marrow cells. One antibody turned out to activate the receptor in a way that induces marrow stem cells -- which normally develop into white blood cells -- to become neural progenitor cells, a type of almost-mature brain cell.
Nature's Toolkit
Natural antibodies are large, Y-shaped proteins produced by immune cells. Collectively, they are diverse enough to recognize about 100 billion distinct shapes on viruses, bacteria and other targets. Since the 1980s, molecular biologists have known how to produce antibodies in cell cultures in the laboratory. That has allowed them to start using this vast, target-gripping toolkit to make scientific probes, as well as diagnostics and therapies for cancer, arthritis, transplant rejection, viral infections and other diseases.
In the late 1980s, Lerner and his TSRI colleagues helped invent the first techniques for generating large "libraries" of distinct antibodies and swiftly determining which of these could bind to a desired target. The anti-inflammatory antibody Humira®, now one of the world's top-selling drugs, was discovered with the benefit of this technology.
Last year, in a study spearheaded by TSRI Research Associate Hongkai Zhang, Lerner's laboratory devised a new antibody-discovery technique -- in which antibodies are produced in mammalian cells along with receptors or other target molecules of interest. The technique enables researchers to determine rapidly not just which antibodies in a library bind to a given receptor, for example, but also which ones activate the receptor and thereby alter cell function.
Lab Dish in a Cell
For the new study, Lerner laboratory Research Associate Jia Xie and colleagues modified the new technique so that antibody proteins produced in a given cell are physically anchored to the cell's outer membrane, near its target receptors. "Confining an antibody's activity to the cell in which it is produced effectively allows us to use larger antibody libraries and to screen these antibodies more quickly for a specific activity," said Xie. With the improved technique, scientists can sift through a library of tens of millions of antibodies in a few days.
In an early test, Xie used the new method to screen for antibodies that could activate the GCSF receptor, a growth-factor receptor found on bone marrow cells and other cell types. GCSF-mimicking drugs were among the first biotech bestsellers because of their ability to stimulate white blood cell growth -- which counteracts the marrow-suppressing side effect of cancer chemotherapy.
The team soon isolated one antibody type or "clone" that could activate the GCSF receptor and stimulate growth in test cells. The researchers then tested an unanchored, soluble version of this antibody on cultures of bone marrow stem cells from human volunteers. Whereas the GCSF protein, as expected, stimulated such stem cells to proliferate and start maturing towards adult white blood cells, the GCSF-mimicking antibody had a markedly different effect.
"The cells proliferated, but also started becoming long and thin and attaching to the bottom of the dish," remembered Xie.
To Lerner, the cells were reminiscent of neural progenitor cells -- which further tests for neural cell markers confirmed they were.
A New Direction
Changing cells of marrow lineage into cells of neural lineage -- a direct identity switch termed "transdifferentiation" -- just by activating a single receptor is a noteworthy achievement. Scientists do have methods for turning marrow stem cells into other adult cell types, but these methods typically require a radical and risky deprogramming of marrow cells to an embryonic-like stem-cell state, followed by a complex series of molecular nudges toward a given adult cell rate. Relatively few laboratories have reported direct transdifferentiation techniques.
"As far as I know, no one has ever achieved transdifferentiation by using a single protein -- a protein that potentially could be used as a therapeutic," said Lerner.
Current cell-therapy methods typically assume that a patient's cells will be harvested, then reprogrammed and multiplied in a lab dish before being re-introduced into the patient. In principle, according to Lerner, an antibody such as the one they have discovered could be injected directly into the bloodstream of a sick patient. From the bloodstream it would find its way to the marrow, and, for example, convert some marrow stem cells into neural progenitor cells. "Those neural progenitors would infiltrate the brain, find areas of damage and help repair them," he said.
While the researchers still aren't sure why the new antibody has such an odd effect on the GCSF receptor, they suspect it binds the receptor for longer than the natural GCSF protein can achieve, and this lengthier interaction alters the receptor's signaling pattern. Drug-development researchers are increasingly recognizing that subtle differences in the way a cell-surface receptor is bound and activated can result in very different biological effects. That adds complexity to their task, but in principle expands the scope of what they can achieve. "If you can use the same receptor in different ways, then the potential of the genome is bigger," said Lerner.
In addition to Lerner and Xie, contributors to the study, "Autocrine Signaling Based Selection of Combinatorial Antibodies That Transdifferentiate Human Stem Cells," were Hongkai Zhang of the Lerner Laboratory, and Kyungmoo Yea of The Scripps Korea Antibody Institute, Chuncheon-si, Korea.
Funding for the study was provided by The Scripps Korea Antibody Institute and Hongye Innovative Antibody Technologies (HIAT).
Milk teeth are a valuable source of stem cells
Interview with Dr. Alexiev Venelin.
Since when there is a procedure to remove teeth to derive stem cells?
In 2003, American scientists discovered that the pulp of milk teeth is a valuable source of biological mesenchymal stem cells that can be isolated and used cryopreservatеа treatment at a critical moment for the man. Scientific achievement is enormous. It's most popular method of extracting stem cells from the umbilical cord and placenta, add another one to the undeniable advantages. It gives parents a second chance, missed the first - to preserve stem cells at birth of their children. Today technology is successfully practiced in the U.S., UK, Greece and Bulgaria in two years.
How and what are the indications for extraction of milk teeth?
Milk tooth extraction is a safe, natural and completely noninvasive method for the extraction and storage of stem cells. Appropriate age from 5 to 12 years. For starters dentist tooth determine whether appropriate, inspection of front upper and lower teeth. Required tooth is with mild shaking.
It is the root to be fully preserved tooth so not only leaves fall and be removed as soon as it starts to shake. Before the operation is done or sectoral panoramic photograph to determine the condition of the tooth and its removal is performed under local anesthesia.
Remove the tooth is placed into a special set of transportation and transported quickly to the laboratory. The Bank tooth is examined to extract stem cells are stored at -196 C ˚. The entire process is accompanied by protocols to ensure the unique genetic material. Finally, the child's parents receive a certificate for successfully storing an initial period of 20 years.
Experts recommend extraction of two teeth, because the pulp of a tooth leads to storage of biological material a sample application. Medical logic leads to the more material you have, the more therapeutic applications are given.
Which of milk tooth are stem cells?
In milk tooth pulp in the accumulation of dentin formed hermetically sealed and sterile space, which contains multiple stem cells. The pulp of the tooth is formed even in the embryonic stage of development of the organism and therefore the cells are young and are carriers of the original DNA. It has been shown that the pulp of a tooth contains from 1000 to 100 thousand units stem cells that can be isolated to reproduce by cell cultures to be implanted in the area of the lesion, giving rise to a new tissue.
What is the application?
Stem cells from milk tooth is defined as mesenchymal, which have the ability to differentiate into tissue-forming cells - heart muscle, kidney, liver, muscle, tendons, cartilage, have the ability to form dentin. Currently, the treatment of diseases through tissue regeneration by mesenchymal stem cells is the most recent and rapidly developing trend in modern medicine.
Research into stem cell therapy is rapidly evolving and offer hope for the treatment of juvenile diabetes, heart disease, arthritic disease, Parkinson's disease, Alzheimer's, spinal cord injury, multiple sclerosis and others. Japanese scientists have managed to create even new teeth in mice. All this is due to the ability of stem cells to differentiate into other cell types. Stem cells are the first motto of cells formed after fertilization, as these are the foundation of the dental pulp: mesenchymal, chondrocytes, osteoblasts and adipocytes.
The fact is that stem cells isolated from cord blood and placenta are much stronger and more numerous than any other. Since there are immunologically mature, they are able to transform into different types of blood cells, making a real alternative for the treatment of 80 types of diseases, including leukemia, disease and Hodgkin lymphoma, breast cancer and testicular multiple sclerosis , a number of neurological diseases and others.
Compared to undifferentiated cells derived from other tissue stem contained in the pulp of milk teeth, however, are very valuable because they reproduce faster, easier to differentiate into other cell types and can be extracted in many wider time range. But with aging stem cells slow their recovery and become much more efficient.
Therefore scientific theory is that the earlier draw, the more effective they will be in time.
Stem cells at birth are affected by external factors and aging
Dr. Svetla Bogdanova, a specialist neurologist. He graduated in medicine in 1985 at the Medical University of Pleven, Bulgaria. In 1990 acquired specialty of neurology at the Medical University - Sofia, Bulgaria.
Passed courses and specializations in "Thermal burns" to MHATEM "Pirogov", "General EEG" at the Medical University - Sofia neurophysiology, "Doppler Neurosonology" at the Medical University - Sofia "Health Management" at the Medical University - Plovdiv. She has worked as head of the Regional Coordination Headquarters - Center for Emergency Medical Care, Sofia. He is currently a neurologist at Military Medical Academy and Medical Manager of Tissue stem cell bank "Biohelenika."
Really stem cells gives rise to the human body?
For the last 4-5 years in many international forums symposia and publications advocates topic of stem cells. Detailing their meaning and application. Each organ is composed of cells, such as liver is composed of hepatocytes, neurons of the brain, bone osteocytes, blood leukocytes and erythrocytes from lung pulmotsin etc. All these cells are highly specialized and differentiated function they perform. Stem cells are precisely those that develop in the earliest stages of embryonic development and have assigned a function to be performed. This is what is valuable. They have the original genetic material of an individual, not differentiated, ie the future is ahead of them and can be converted (transformed) into specialized cells of various organs. The most important thing about stem cells is that they are born the youngest cells, unaffected by external factors and aging. This is unique.
Is proven and diseases which are used?
Bone marrow transplantation was first performed in 1968 Bone marrow cells are used to treat patients diagnosed with leukemia, aplastic anemia, lymphoma, Hodgkin's disease, immune disorders, cancer. We know how is hard to find a suitable donor, how long to wait and how costly it. The decision to proceed with private banking (storage) of stem cells, so is the business organization and authorization of Biohelenika - stem cell bank, of which I am medical director. Modern medicine has come to the conclusion that everyone is good to have available their own stem cells.
Since 1988, stem cells from umbilical cord was successfully applied to replace the bone marrow. Stem cell transplants allow treatment of many serious diseases. This type of therapy have proven their undeniable advantage in the treatment of blood cancers - leukemia and lymphoma, is increasing. 107.3 to 100 thousand in 2005
Scientists create teeth with implants grown from stem cells
We want it or not, teeth wear out with age comes a time when you need to replace them with artificial ones. But now there may be an alternative to artificial prostheses - and it is real teeth, "grown" in a lab and created by human cells wreath.
Studies were conducted with mouse stem cells are derived from embryos and specially selected so as to be able to produce growth in other cells to grow out of their teeth. So the combination of murine stem cells and human gingival cells in the laboratory has produced a real tooth - shaped with enamel and roots. The roots were fully alive, suggesting that transplanted into human mouth, most likely belonged to the bone.
The results of this discovery may mean turning point in the treatment of periodontal disease and produce new teeth in people who for some reason have lost their. Of course, the stem cells in the formation of teeth, suitable for implantation in humans must be human rather than mouse and extracting stem cells from human embryos for scientific purposes is not perceived as ethical. Scientists argue that stem cells can be easily extracted from the pulp of human wisdom as to produce the same effect.
Retrovirus Packaging
We provide high quality Retrovirus packing services.
Retroviral vectors are one of the efficient methods of gene delivery to mammalian cells both in vitro and in vivo. Through years of experience with lentiviral and retroviral vectors, Targatt has developed own proprietary packaging systems and efficient protocols for the rapid generation of pseudoviral particles.
Targatt offers express Retrovirus packaging service to produce high-quality, high-titer virus particles using your viral construct with turnaround time of 10 days. Save your time, receive ready-to-transduce viral particles.
- Production of virus in a state-of-the art BSL-2 facility with robust quality control in accordance with NIH Biosafety Level 2 criteria
- Flexibility of virus production scales to meet your research needs
- Accurate Virus titers as determined by qPCR to measure infectious units per ml ( ifus/ml)
- Depending on level of titer requested, please provide 10-40 μg of endotoxin-free lentivector or retrovector plasmid DNA
- All custom virus production services should come with information on the plasmids which have to be packaged.
- Vector must be able to produce virus (no internal poly A signal, no toxic genes, no unusual 2nd structure, or 5' to 3' LTR size over 8kb.)
Lentivirus Packaging
Targatt provides high quality and high titer 10-day Lentivirus packing services.
Lentiviral / retroviral vectors are one of the most efficient methods of gene delivery to mammalian cells both in vitro and in vivo. Through years of experience with lentiviral and retroviral vectors, Targatt has developed own proprietary packaging systems and efficient protocols for the rapid generation of pseudoviral particles.
- offers express lentiviral/retrovirus packaging service to produce high-quality, high-titer virus particles using your viral construct with turnaround time of 10 days. Save your time and receive ready-to-transduce viral particles.
- Production of virus in a state-of-the art BSL-2 facility with robust quality control in accordance with NIH Biosafety Level 2 criteria
- Flexibility of virus production scales to meet your research needs
- Accurate Virus titers as determined by qPCR to measure infectious units per ml ( ifus/ml)
- Depending on level of titer requested, please provide 10-40 μg of endotoxin-free lentivector or retrovector plasmid DNA
- All custom virus production services should come with information on the plasmids which have to be packaged.
- Vector must be able to produce virus (no internal poly A signal, no toxic genes, no unusual 2nd structure, or 5' to 3' LTR size over 8kb.)
Stem Cell Culture Services
We offer standard and customized cell culture services including mycoplasma testing and FBS lot evaluation.
Please contact us to discuss your needs with our technical service specialists.
Stem Cell Differentiation
Targatt offers various services for the differentiation of ESC/iPSC lines into more specialized cells including multipotent stem cells and fully differentiated somatic cells.
Advantage of using multipotent stem cells
- Thaw ESC/iPSC-derived multipotent stem cells (e.g. Neural Stem Cells) when you need them
- Differentiate multipotent stem cells to specialized somatic cells (e.g. neurons and glial cells)
- Expand the multipotent stem cells for a ready supply for your planned experiments = save money and time!
Standard services:
- Neural Stem Cells
- Neural Crest Stem Cells
- Cardiomyocytes
- Skeletal Muscle
Bespoke services:
We also offer customized differentiation services. Please contact us with details about what kind of cells you are interested in.
Human ESC / iPSC Neuronal Differentiation Service
- Send us your pluripotent cell line (or use our iPSC generation service)
- We will differentiate your ESC/iPSC line into neural stem cells (NSCs) or neural crest stem cells (NCSC) by using our proprietary neural induction protocol.
- NSCs and NCSCs are proliferating cells that can be frozen and cultured over a prolonged period of time.
- NSCs can be further differentiated into multiple neural cell types.
Stem Cell Characterization
Human and Mouse ESC/iPSC Characterization in vivo & in vitro
- Teratoma Formation Analysis Service
- EB Formation and Analysis Service
- Pluripotency Marker Characterization (Human / Mouse)
- Karyotyping (Human / Mouse)
- Germline Transmission of mouse ESCs
- Sex determination
We make the line for you, and we characterize it, too!
We offer a wide range of services to help you identify and characterize stem cell lines.
Stem Cell Generation
We have successfully derived many induced pluripotent stem cell (iPSC) lines from patient specific fibroblasts.
We have three methods for iPSC generation: mRNA, episomal (both genomic footprint-free), or retroviral.
Genomic footprint-free Induced Pluripotent Stem Cell (IPSC) Generation Service:
- - No genomic footprint - the non-integrating system is safe in drug discovery and cell therapy applications.
- - Two methods available: Episomal DNA vectors containing the reprogramming factors or mRNA.
- - High reprogramming efficiency (0.2%-0.5%).
- - The reprogramming vectors/mRNA are removed naturally during the cell cycle.
- - Safe to handle - no viral particles.