At the beginning of the month, the California Institute for Regenerative Medicine (CIRM) announced its first Tools and Technologies Awards. Established in 2004 as the result of the passage of a California ballot proposition, the nonprofit CIRM is charged with distributing $3 billion in loans and grants for stem cell research to public and private entities in the state. The CIRM has thus far awarded 253 grants worth more $635 million. Eighteen two-year grants totaling more than $19 million were awarded to 23 institutions for the creation or scale-up of new techniques for stem cell research and therapy (see table, page 6). Of the 18 grants, six went to private companies, which received a total of nearly $6 million in funding. This funding will be used to create new instruments, reagents, stem cell lines and disease models that could eventually be commercialized.

Reagent and software firm Vala Sciences received a grant to produce and study human embryonic stem cell (hESC)-derived cardiomyoctyes (CM), which could potentially enable myocardial repair and drug development. HESCs are pluripotent, in that they have the ability to grow into any type of cell, including CMs, a heart muscle cell. Damage to CMs is responsible for heart failure and heart disease, among other ailments.

Vala plans to generate antibiotic-resistance hESC lines (antibiotic resistance is a type of marker for a desired cell trait) in order to isolate and study hESC-CMs. “[The grant is for] improving CM subtype identification, selection and induced maturation of subtypes of CM from immature and primitive stem and progenitor cell-derived CM,” said Casey Laris, Vala’s director of marketing.

To study CM differentiation (how CMs develop into specific CM subtypes) Vala will employ its automated fluorescence image cytometry technique (developed with the Burnham Institute for Medical Research). “The company’s imaging technology is derived from very high-content screening technology. It is basically an automated microscope that we’ve then applied algorithms to automate cell by cell analysis to differentiate cell populations,” stated Mr. Laris. Specific areas of interest include the role of cardiac-specific promoters in CM-subtype development, the electrophysiological properties of CM subtypes and CMs’ interactions with extra-cardiac cells. The goal of the project, as stated in the company’s grant application, is to develop an in vitro system to optimize the selection of CM phenotypes.

Gamma Medica-Ideas’s grant will be used to produce a prototype instrument that is capable of 3-D imaging of single stem cells within live animals. The instrument will combine magnetic resonance imaging (MRI) and single photon emission computer tomography (SPECT) technologies to provide live-animal interior images of up to 1 cm3. According to Gamma’s grant application, current technology can image up to 0.3 mm within live animals. A combination system would allow researchers to combine the information about tissue and cellular changes that SPECT provides with the anatomical images of MRI in order to determine the position of cells in relation to identifiable sites within an animal.

Due to size of the vacuum tube technology used by traditional gamma cameras, there are no combination SPECT-MRI systems on the market, according to Gamma. At the end of 2008, Gamma Medica released the FLEX Pre-Clinical Imaging Platform, which replaced the standard SPECT vacuum tube technology with a 50 µm-resolution Megapixel camera. With the grant money, Gamma Medica will construct a high-resolution, low-energy SPECT system that will be placed within an MRI to provide single-cell imaging. Gamma Medica-Ideas received a $100,000 grant from the National Institutes of Health in fall of 2006 for a SPECT/MRI imaging instrument.

The CIRM grant jointly awarded to Fluidigm and Stemgent will fund the creation of a microfluidic chip to study the proper conditions for stem cell differentiation, as well as the characterization and identification of the genes and small molecules involved in stem cell differentiation. The product prototype is designed to be available for manufacture upon completion. According to the CIRM’s review summary of the application, the project will focus on the reprogramming of somatic cells (all human cells excluding reproductive cells) to induce pluripotent stem cells.

In 2007, multiple studies found that it is possible to convert differentiated cells (cells specialized for a particular function) into pluripotent stem cells. Fluidigm will redesign its current integrated fluidic circuit technology to accommodate stem cell culturing. According to Fluidigm, the chambers of the microfludic chip will culture specific cells, which will tested against various with reagents. Results will be analyzed using light and fluorescent microscopy. Stemgent, a company that specializes in reagents for stem cell research, will develop reagents for the project. The CIRM states in its review summary that the project will utilize 16 reagents.

The grant also advances another study published last year. Researcher at Stanford University and the University of Pennsylvania designed an automatic microfluidic chip-based cell culture-screening system with 96 wells. The purpose of the experiment was to determine if an automatic microfluidic chip was able to contain and differentiate stem cell cultures for sustained experiments. According to the study, the advantages of a microfluidic chip for this research are its ability to provide microscopic information, while patterning the cell culture substrates, allowing researcher to have more control over the conditions of the cells. GE Advanced Materials fabricated the chip used in the experiment by employing multiplayer soft lithography. Fluidigm’s use of multiplayer soft lithography was cited by CIRM reviewers as one of the reasons behind the grant’s approval and Fluidigm cites the research in its announcement of the grant..

Invitrogen, part of Life Technologies, received a grant to use homologous recombination (HR), a technique for genetic modification, to develop in vitro models of Lou Gherig’s disease (ALS) in hESCs. Invitrogen plans to create hESC lines with genetic mutations similar to those of ALS patients, including specific cell types. As a result, Invitrogen expects to develop a protocol for HR in hESCs, as well as related cell reporter genes and hESC cell lines. In speaking to IBO, a spokesperson from Invitrogen stated that the researchers have not decided on the transfection reagents to be used in this project. Current transfection reagent product lines produced by Invitrogen for use in stem cells include Lipofectamine 2000, ViraPower Lentiviral Technology and the ViraPower Promoterless Lentiviral Gateway Expression System.

VistaGen Therapeutics, a developer of cell-based assays, received a grant to create a technique to help produce more mature hESC-derived liver cells. Such cells could serve as models for drug metabolism and thus for drug screening. According to VistaGen, the current protocols for producing hESC-derived liver cells result in immature cells. VistaGen proposes to develop a tool that will tag a protein found in mature liver cells, enabling an improve ways to create mature hESC-derived liver cells. VistaGen plans to license the technology, including associated reagents.

Stem cell–engineering firm Novocell received a grant to further develop a implantable, retrievable device for the delivery of insulin-producing cells in mice. The device would allow for the implant without the use of immunosuppressant drugs. According to its grant application, Novocell will study the device’s use with hESC-dervied pancreatic progenitor cells, which are more limited in their differentiation than stem cells. Novocell received a research grant from the CIRM earlier this year.