Scalable protocol to differentiate skeletal muscle cells

AMSBIO announces the availability of the world's first kit to differentiate human pluripotent stem cells into functional myotubes. The new kit utilises a highly efficient media based protocol to produce skeletal muscle cells from stem cells in a simple, scalable manner.

The potential to differentiate stem cells into specific cell types is revolutionising life sciences, from new methods of studying developmental biology and novel approaches to producing accurate disease models to techniques to help drug discovery and toxicity testing.

Until recently methods of studying muscular disease and potential therapies were dependent on invasive muscle biopsies to produce limited batches of primary cells. Use of primary cells presents challenges, not only in the collection process but also related to inconsistencies in cell growth, behaviour and life span, making it difficult to generate reliable experimental models.

The new Skeletal Muscle Differentiation kit offers researchers a unique tool to rapidly differentiate donor stems into functional myotubes in a reproducible fashion. Tested on a wide range of human embryonic and induced pluripotent stem cell lines the new kit follows a simple three-step process of media changes and cell passaging. Eliminating the need for cell sorting or transfection of myogenic transcription factors, the three step protocol generates a highly pure population of approximately 70% skeletal muscle myotubes in weeks.

Simultaneous 96-well extraction of urine samples for drugs of abuse analysis

INTEGRA, in conjunction with DPX Labs LLC, has produced an application note that describes a novel high throughput semi-automated sample method for processing urine samples ready for drugs of abuse analysis by LC-MS/MS.

Using an INTEGRA VIAFLO 96 electronic pipette with DPX mixed mode tips, a single 96-well plate loaded with samples can be extracted and ready for LC-MS/MS analysis in less than 10 minutes eliminating the need for a time-consuming evaporation step.

Sample preparation is required to remove matrix interferences from urine samples prior to LC-MS/MS analysis. This procedure is typically very time-consuming and is generally the “bottleneck” for laboratory analysis. DPX extraction is a highly reproducible and sensitive dispersive SPE method that requires much less solvent compared to other SPE techniques.

The application note describes how an INTEGRA VIAFLO 96 electronic pipette was used to simultaneously undertake, in each well of a 96-well plate, the various steps of the DPX protocol (analyte binding, resin washing, analyte elution). Results from this semi-automated method are shown to be linear, accurate, and reproducible.  All correlation coefficients for the protocol were greater than 0.99 for the range of 12.5–400 ng/mL.

The INTEGRA VIAFLO 96 is a handheld 96-channel electronic pipette that enables fast, precise and easy simultaneous transfer of 96 samples from microplates without the cost of a fully automated system. The VIAFLO 96 requires no special skills or training to operate it. Fast replication or reformatting of 96-well plates and high precision transferring of reagents, compounds and solutions to or from microplates with the VIAFLO 96 is as easy as pipetting with a standard electronic pipette into a single tube. Four pipetting heads with pipetting volumes up to 12.5µl, 125µl, 300µl or 1250µl are available for the VIAFLO 96. These pipetting heads are interchangeable within seconds enabling optimal matching of the available volume range to the application performed. 

Non-invasive online monitoring of biomass in shake flasks

aquila biolabs’ Cell Growth Quantifier (CGQ) allows non-invasive online monitoring of biomass in shake flasks. The automatic measurements save time and provide a profound understanding of the bioprocesses, ensuring sustainable success of shake flask experiments.

Currently, scientists monitor biomass in shake flask cultures by manual and invasive photometric OD measurements. The CGQ, however, determines the biomass concentration automatically, optically and non-invasively through the vessel wall using a patented sensor. Overall, the CGQ can generate detailed microbial growth kinetics in up to 16 shake flasks simultaneously and in real time.

The benefits are obvious: time and cost savings, an undisturbed bioprocess and improved reproducibility of results. Thanks to the CGQ, bioprocesses can be developed and optimized based on detailed growth curves. Hence, the sustainable success of shake flask experiments can be significantly increased.

The CGQ consists of four components. The sensor plate is positioned under the shake flask, which is protected by a cover. By connecting the shake flasks to the base station, an interface to the CGQuant software is established, which analyses and visualises the biomass data.

The CGQ is compatible with all INFORS HT shakers and clamps as well as with 'Sticky Stuff'. All common types of glass and transparent single use flasks in the sizes 250, 300, 500, 1000 and 2000ml can be used. Thereby, customers can easily integrate the CGQ into their existing laboratory infrastructure.

Automating blood and stem cell banking

Reliance Precision’ s Cool Muscle motors have now been used in 500 Biosafe Sepax systems. The Sepax systems are used for automated adult stem cell banking and cell processing applications.

The Biosafe Sepax system is an automated, mobile, closed unit for the efficient and consistent processing of umbilical cord blood, bone marrow, peripheral blood or other cell based products. Biosafe began trialling Reliance’s Cool Muscle motors in 2010 in its Sepax systems, using the motor to help accurately separate the different cell components via single-use cell separation kits.

The Cool Muscle motor senses when the different single-use cryobags are filled with the correct amount of cellular products and was so successful in trials that Biosafe continued to use it in its Sepax systems, more than 500 of which are now in use worldwide.

Julien Camisani, Scientific and Technical Director at Biosafe, explains: “We used to have step motors in our automated blood banking systems but this process was sometimes inefficient. Since adding the Cool Muscle motors to our Sepax systems we have not experienced any of the issues we did when using step motors. We are so satisfied with the excellent quality of the Cool Muscle motors that we intend to continue using them in our Sepax systems and in future development projects.”

New ChIP-seq protocol yields improved results

Chroma trap has developed a new protocol that further extends the many advantages of its popular Chromatin Immunoprecipitation Sequencing (ChIP-seq) assay kits.

Key developments for the Chromatrap ChIP-seq kit (version 1.2) include high quality chromatin can now be achieved via sonication or enzymatic digestion; high and low abundant enrichment is possible from small chromatin samples; improved antibody binding; greater chromatin loading flexibility and the ability to use increased slurry volumes for difficult samples.

Chromatrap’s ChIP-seq kit uses solid state technology in parallel with high throughput sequencing to deliver a streamlined ChIP-seq protocol from small cell numbers and low chromatin concentrations. Specifically adapted for broader chromatin concentrations, Chromatrap ChIP-seq combines the dynamic range of Chromatrap with the downstream analysis power of deep sequencing. This allows faster, more reproducible genome wide identification of TF binding sites and specific DNA associated protein modifications. With no limitation in scale or resolution, Next Generation Sequencing can elucidate the role of TFs and epigenetic marks on gene transcription and epigenetic chromatin status.

The Chromatrap ChIP-seq kit allows the user to perform up to 24 ChIP assays from cell collection through to immunoprecipitation, including up to 10 chromatin sample preparations. The kit provides all of the major components required for performing ChIP assays to obtain high quality DNA for Next Generation Sequencing library preparation.

With the Chromatrap ChIP-seq kit you can sequence from as little as 1-50 µg of chromatin and perform up to 10 library preparations form a single IP. A complete ChIP-seq assay can be completed in just five days. With selective and sensitive enrichment of low chromatin loading and optimised elution buffer chemistry the new version 1.2 of the Chromatrap’s ChIP-seq kit enables preparation of high quality and quantity of immunoprecipitated DNA.