Above: hESC in ESC-Sure
conditioned medium
Bottom: hESC on MEF TM

Background: Traditionally human embryonic stem cell (hESC) culture requires a mouse embryonic fibroblast (MEF) cells’ support, which makes the protocol complicated. Our Serum- and Feeder-free medium that is for human ESC culture contains all the growth factors needed in hESC culture, except the bFGF; eliminate the routine preparation of feeder. It is a ready-to-use product for a serum/feeder-free system. Our medium makes your cell culture more efficient and easier to control.
Serum- and Feeder-free medium (1x) for proliferation of pluripotent human embryonic stem cell (hESC) culture in a serum/feeder-free system. Each and every batch has been tested for hES/iPS cells pluripotency. All you need to add is 20 ng/ml bFGF.
Product description: 100 mL of hESC-Sure
Source: Chemical plus growth factors
Storage: -80°C
Shipping: Shipped on dry ice.
Recommended procedure:
Human ESC culture procedure using hESC-Sure Serum- and Feeder-free medium:
1. Coat culture dish with Matrigel the day before.
2. Wash Matrigel (BD Biosciences Cat# 354277) coat dishes with DMEM/F12 medium(Targatt Cat# ASM-5002).
3. Once human ES cell were confluent or cultured for more than 5 days, disaggregated cells
with 1mg/ml Dispase(Dilute 5mg/ml stock solution with DMEM/F12 media), incubate at 37
C for 3 minutes.
4. Wash container with 1x PBS, and to scrape colonies off the bottom of the container with cell
scraper.
5. Collect the cells in the 50ml or 15ml falcon tube, pellet by centrifugation at 700rpm for 5 min
at room temperature.
6. Resuspend hESC pellets in hESC-Sure Serum- and Feeder-free culture medium supplemented with 20 ng/ml bFGF.
TM
7. Split cells at 1:3 to 1:5 ratio every 4 to 5 days using 1mg/ml Dispase (diluted with
DMEM/F12).
8. 1x freezing medium contains 90% knockout serum and 10% DMSO.

 Price: 104 EUR

Gene knock-down using short hairpin RNA (shRNA) to inhibit expression of the corresponding gene has been widely used in gene function studies, drug discovery, and disease research.  Our TARGATT Technology allows us to generate in vivo shRNA knock-down mouse models in 3 months.

Targatt can create site-specific knock-in mice for you within 3 months. Using our novel TARGATT system, a gene of interest can be inserted at a well-characterized, transcriptionally-active locus in the mouse genome with guaranteed transgene expression. Tissue-specific and/or ubiquitous expression options are available. Please contact us for a list of plasmid construct with reporter genes. 

In addition to our full service we also offer individual service options to generate your TARGATT knock-in mouse.

You can prepare the DNA using the TARGATT Kit and we do the pronuclear microinjection for you. Or we can prepare the DNA that you then inject after setting up your own TARGATT mouse colony.

Please contact us to discuss your project plan for your fast & site-specific knock-in or TARGATT Knock-down Mouse services.

Un bebé de dos años que nació con el VIH ha sanado con terapia antirretroviral precoz

Ayer, investigadores de amfAR, (Fundación para la investigación del sida en Estados Unidos) hicieron público en el marco de la 2013 Conference on Retroviruses and Opportunistic Infections, el primer caso documentado de curación de sida en un bebé nacido con VIH. Esta sanación, que fue posible con terapia precoz de antirretrovirales según los médicos, señala la necesidad de investigar más sobre el efecto de estos tratamientos en recién nacidos.

Deborah Persaud, doctora de la Johns Hopkins University de Estados Unidos, describió ayer, tres de marzo de 2013, el primer caso documentado de un bebé curado del sida. Su anuncio fue realizado en la 2013 Conference on Retroviruses and Opportunistic Infections (CROI) de Atlanta (EEUU). 

Persaud, investigadora de amfAR (The Foundation for AIDS Research) , detalló el caso de un bebé de dos años de edad de Mississippi diagnosticado con VIH al nacer, y que enseguida fue sometido a terapia antirretroviral. 

A los 18 meses, el bebé dejó de tomar los antirretrovirales y su seguimiento médico se interrumpió. Cuando los médicos volvieron a verlo a los 23 meses, a pesar de que había dejado de su terapia durante cinco meses, constataron que el bebé tenía una carga viral indetectable. Una batería de pruebas altamente sensibles posteriores confirmó la ausencia del VIH. 

La confirmación de la curación fue posible gracias a una donación que amfAR concedió a la doctora Persaud y a la doctora Katherine Luzuriaga, de la Universidad de Massachusetts, en septiembre de 2012. 

La donación permitió que las médicos establecieran un colaboratorio de investigación para explorar y documentar posibles casos pediátricos de curación del VIH. En este colaboratorio participan además los doctores e investigadores Stephen Spector y Richman Doug, de la Universidad de California, San Diego; el Dr. Frank Maldarelli, del Instituto Nacional del Cáncer, y el Dr. Tae-Wook Chun, del Instituto Nacional de Alergias y Enfermedades Infecciosas.

Diversas formas de tratamiento del VIH

"El pediatra del bebé de Mississippi conocía nuestro trabajo y comunicó el caso a nuestro equipo, tan pronto como se enteró," explica Rowena Johnston, vicepresidenta de amfAR. "Gracias a que este colaboratorio ya estaba en marcha, los investigadores pudieron movilizarse inmediatamente y realizar las pruebas necesarias para determinar si realmente se trataba de un caso de curación del SIDA infantil”. 

Según Persaud, pruebas exhaustivas han confirmado sin lugar a dudas que la madre y el bebé eran VIH positivo cuando este nació, y que en la actualidad no quedan signos de infección por VIH en el bebé, según los análisis realizados con los medios más sensibles disponibles. 

El único caso documentado de cura del VIH hasta la fecha había sido el de Timothy Brown, el llamado "paciente de Berlín." En 2006, mientras era tratado por el VIH, al Sr. Brown le diagnosticaron leucemia. 

Su médico trató entonces su leucemia con un trasplante de células madre de una persona nacida con una mutación genética que causa la inmunidad a la infección por VIH. Después del trasplante, el Sr. Brown pudo abandonar el tratamiento para el VIH sin recaídas. 

Este nuevo caso apunta a la posibilidad de que diferentes poblaciones de personas con VIH podrían ser curadas de esta enfermedad de diferentes maneras. Mientras que el caso del señor Brown fue el resultado de una serie de complejos y costoso procedimientos de alto riesgo, este nuevo caso parece haber sido el resultado directo de una terapia antirretroviral relativamente barata. 

"Teniendo en cuenta que esta sanación parece haberse logrado solo con terapia antirretroviral, es imperativo que aprendamos más acerca del sistema inmunológico de los recién nacidos y en qué se diferencia este del sistema inmunológico de los adultos; así como sobre los factores que han hecho posible que el bebé se haya curado”, explica Johnston. 

El caso de Mississippi también pone de relieve la importancia de la identificación del VIH en mujeres embarazadas y de ampliar el acceso a tratamientos para prevenir la transmisión materno-infantil de la enfermedad. Asimismo, revela la necesidad de tratar de inmediato con antirretrovirales a los bebés que nacen seropositivos. 

Acerca de amfAR

amfAR, la Fundación para la Investigación del SIDA, es una de las principales organizaciones sin fines de lucro del mundo dedicada al apoyo de la investigación del SIDA, prevención del VIH, educación, tratamiento y su incidencia en las políticas públicas. Desde 1985, amfAR ha invertido más de 366 millones de dólares (unos 280 millones de euros) en sus programas y ha otorgado becas a más de 2.000 equipos de investigación de todo el mundo.

TARGATT Transgenic Kit is designed to create site-specific, knock-in transgenic mice in a more efficient and significantly faster way over traditional methods. Generating transgenic mice by conventional methods (e.g. pronuclear microinjection or lentiviral injection) has several limitations, first of which is random insertion of the transgene. Random insertion of a transgene results in a position effect where either the transgene is prone to silencing or endogenous gene expression is disrupted. Furthermore, this position effect is compounded when a transgene is inserted as multiple copies, resulting in instability at the insertion locus.

Using our proprietary site-specific DNA integration system, TARGATT Transgenic Kit, combined with our genetically TARGATT mouse embryos (cat.# AST-0001, AST-0002, AST-0003, AST-0004). Targatt offers high quality H11(Hipp11) and ROSA26 transgenic mouse kits with our TARGATT technology. Using our TARGATT Technology, you can generate your own knock-in mouse in just three months.

TARGATT embryos are derived from one of our genetically engineered mouse models. These mouse embryos can be used as embryo donors for creating site-specific transgenic mice in a more efficient and faster way compared to traditional methods. Generating transgenic mice by conventional methods (e.g. pronuclear microinjection or lentiviral injection) has several limitations, first of which is the random insertion of the transgene. Random insertion of a transgene results in a position effect where either the transgene is prone to silencing or endogenous gene expression is disrupted. Furthermore, this position effect is compounded when transgenes are inserted in multiple copies, resulting in genomic instability at the insertion locus. Using our proprietary site-specific DNA integration technology, TARGATT embryos combined with our TARGATT Transgenic Kit (cat# AST-1001 or AST-1002), you can generate your desired transgenic mouse models with guaranteed gene expression faster.

Gentaur offers high quality H11 and ROSA26 TARGATT frozen embryos. Using our TARGATT Technology, you can generate your own knock-in mouse from H11 or ROSA26 embryos in 3 months. Our H11(Hipp11) and ROSA26 frozen embryos contain 3 straws with a total 45-60 cryopreserved embryos (8-cell stage/morula). They are shipped in a dry shipper containing liquid nitrogen. We also offer fresh H11 and ROSA26 embryos with each vial containing 25-35 fresh embryos (E3.5) in KSOM medium. We offer overnight shipping at ambient temperature. Embryos will be transferred to recipient(s) immediately upon arrival. US customers only.

TARGATT Embryos

Using our novel TARGATT system, a gene of interest can be specifically inserted at a well-characterized, transcriptionally-active locus in the mouse genome with guaranteed transgene expression. Tissue-specific and / or ubiquitous expression options are available.

Advantages of TARGATT technology:

1. Site-specific gene integration at a transcriptionally-active locus ensures high-level gene expression.
2. Integration happens at an intergenic region; no internal genes are disrupted.
3. The integrase system catalyzes a unidirectional integration event and results in a high efficiency in producing transgenic mice.
4. Gene integration at the same locus allows a precise comparison of the transgenics from one line to another.

TARGATT Kits

TARGATT Supporting Materials

Mouse Embryonic Fibroblasts (MEF)

●  CF-1
●  Neo-resistant
●  DR4
●  SNL (STO feeder cells)

SNL (STO feeder cells)

Cell Culture Products

●  Germline-tested & ESC-qualified FBS
●  Specialty Media
●  Basal Media (DMEM)
●  Stem Cell Growth and Differentiation Factors
●  ASC Small Molecules
●  3D Culture and Expansion System

Reprogramming

ESC/iPSC Characterization

  Pluripotency Protein Markers                   Stem Cell Gene Array
     

●  Pluripotency mRNA Markers
●  Components

ESC/iPSC Differentiation

●  Neural Differentiation
●  Dendritic Cell (DC) Generation

ES/iPS Cell Lines

Mouse ES Cell Lines                                 Human iPS Cells

Cell Depository

Primary Cell, cDNA, RNA (Normal)

●  Endocrine
●  Neural
●  Pulmonary
●  Digestive
●  Urinary
●  Reproductive
●  Skeletal Muscle
●  Hematopoietic
●  Integumentary
●  Cardiovascular
●  Miscellaneous

Primary Cell, cDNA, RNA (Disease)

●  Blood Disorders
●  Neurological Disorders
●  Degenerative Disorders
●  Metabolic Disorders
●  Cardiovascular Disorders
●  Congenital Disorders
●  Endocrine Disorders
●  Autoimmune Disorders
●  Genetic Disorders
●  Muscular Disorders
●  Oncogenic Disorders

All products - Full view result 

Read more about Targatt gene modification here

Dr. Deborah Persaud of Johns Hopkins University today described the first documented case of a child being cured of HIV. The landmark findings were announced at the 2013 Conference on Retroviruses and Opportunistic Infections in Atlanta, GA.

Dr. Persaud, an amfAR grantee, detailed the case of a two-year-old child in Mississippi diagnosed with HIV at birth and immediately put on antiretroviral therapy. At 18 months, the child ceased taking antiretrovirals and was lost to follow-up. When brought back into care at 23 months, despite being off treatment for five months, the child was found to have an undetectable viral load. A battery of subsequent highly sensitive tests confirmed the absence of HIV.

Confirmation of the cure was made possible by a grant the Foundation awarded to Dr. Persaud and Dr. Katherine Luzuriaga of the University of Massachusetts in September 2012. The grant allowed Drs. Persaud and Luzuriaga to establish a research collaboratory to explore and document possible pediatric HIV cure cases. The collaboratory includes renowned researchers Drs. Stephen Spector and Doug Richman at the University of California, San Diego; Dr. Frank Maldarelli at the National Cancer Institute; and Dr. Tae-Wook Chun at the National Institute of Allergy and Infectious Diseases.

"The child's pediatrician in Mississippi [Dr. Hannah Gay, a pediatric HIV specialist at the University of Mississippi] was aware of the work we were doing, and quickly notified our team as soon as this young patient's case came to her attention," said Dr. Rowena Johnston, amfAR vice president and director of research. "Because the collaboratory was already in place, the researchers were able to mobilize immediately and perform the tests necessary to determine if this was in fact a case of a child being cured."

According to Dr. Persaud, comprehensive tests have confirmed beyond doubt that both mother and child were HIV positive when the child was born, and today no signs of HIV infection in the child can be detected by the most sensitive means available.

The only other documented case of an HIV cure to date remains that of Timothy Brown, the so-called "Berlin patient." In 2006, while on treatment for HIV, Mr. Brown was diagnosed with leukemia. His physician was able to treat his leukemia with a stem-cell transplant from a person who was born with a genetic mutation causing immunity to HIV infection. Following the transplant, Mr. Brown was able to stop HIV treatment without experiencing a return of his HIV disease.

This new case points to the tantalizing possibility that different populations of HIV-positive people might be cured in different ways. While Mr. Brown's case was the outcome of a complex, high-risk, and expensive series of procedures, this new case appears to have been the direct result of a comparatively inexpensive course of antiretroviral therapy.

"Given that this cure appears to have been achieved by antiretroviral therapy alone," said Dr. Johnston, "it is also imperative that we learn more about a newborn's immune system, how it differs from an adult's, and what factors made it possible for the child to be cured."

The Mississippi case also underscores the importance of identifying HIV-positive pregnant women, expanding access to treatment regimens than can prevent mother-to-child transmission, and of immediately putting infants on antiretroviral therapy in the event that they are born HIV positive.

"We are proud to have played a leading role in bringing this first pediatric HIV cure to light," said amfAR CEO Kevin Robert Frost. "The case is a startling reminder that a cure for HIV could come in ways we never anticipated, and we hope this is the first of many children cured of HIV in the months and years to come." 

The AccuPower DNA Ligation PreMix is a lyophilized master mix containing T4 DNA Ligase, ATP, reaction buffer, and patented stabilizer. This DNA ligation premix is conveniently aliquoted in strip-tubes for reactions; you need only add DNAs to be ligated and water. The reaction will work for DNA ligation for all applications: blunt cloning, sticky end cloning and TA cloning. The premix is stable up to four months at room temperature and for three years at -20°C.

Features and Benefits

AccuPower DualStar qPCR PreMix is a lyophilized PCR master mix containing a thermostable DNA polymerase, thermostable pyrophosphatase, reaction buffer, dNTPs, tracking dye, and patented stabilizer in a ready to use hotstart qPCR master mix.

DualStar qPCR PreMix eliminates nonspecific reaction, while providing high sensitivity, due to our novel HotStart methodology. Gentaur uses a unique enzyme-mediated HotStart PCR that provides robust and reproducible results. Gentaur’s Top DNA Polymerase is inhibited at lower temperatures (< 70°C) by pyrophosphate. However, Top DNA Polymerase is rendered fully active at temperatures above 70°C via pyrophosphate hydrolysis with our thermostable pyrophosphatase. This prevents the formation of mis-primed products and primer-dimers during the reaction set up process resulting in improved PCR specificity. It is ideal for nucleic acid amplification reactions involving complex genomic or cDNA templates, very low copy targets, and multiplex reactions.

DualStar Plus qPCR PreMix goes a step further by providing resistance to common inhibitors of PCR (Blood-EDTA, Hemoglobin, Humic Acid from various sources, etc.) providing you with robust and reliable results, even with poor quality samples. Tested for resistance against many common inhibitors of PCR, DualStar plus is the enzyme of choice for any PCR that you want to work right – the first time and every time!

Features and Benefits