Tumor-infiltrating lymphocytes: Streamlining a complex manufacturing process.

Adoptive cell therapy of tumor-infiltrating lymphocytes has shown promise for treatment of refractory melanoma and other solid malignancies; however, challenges to manufacturing have limited its widespread use. Traditional manufacturing efforts were lengthy, cumbersome and used open culture systems. We describe changes in testing and manufacturing that decreased the process cycle time, enhanced the robustness of critical quality attribute testing and facilitated a functionally closed system. These changes have enabled export of the manufacturing process to support multi-center clinical trials.

Novel approach for endothelializing vascular devices: understanding and exploiting elastin-endothelial interactions.

Elastin is an essential component of arteries which provides structural integrity and instructs smooth muscle cells to adopt a quiescent state. Despite interaction of endothelial cells with elastin in the internal elastic lamina, the potential for exploiting this interaction therapeutically has not been explored in detail. In this study, we show that tropoelastin (a precursor of elastin) stimulates endothelial cell migration and adhesion more than smooth muscle cells. The biological activity of tropoelastin on endothelial cells is contained in the VGVAPG domain and in the carboxy-terminal 17-amino acids. We show that the effects of the carboxy-terminal 17 amino acids, but not those of VGVAPG, are mediated by integrin α(V)ß(3). We demonstrate that tropoelastin covalently linked to stainless steel disks promotes adhesion of endothelial progenitor cells and endothelial cells to the metal surfaces. The adherent cells on the tropoelastin-coated metal surfaces form monolayers that can withstand and respond to arterial shear stress. Because of the unique effects of tropoelastin on endothelial and smooth muscle cells, coating intravascular devices with tropoelastin may stimulate their endothelialization, inhibit smooth muscle hyperplasia, and improve device performance.

Human glial-restricted progenitor transplantation into cervical spinal cord of the SOD1 mouse model of ALS.

Cellular abnormalities are not limited to motor neurons in amyotrophic lateral sclerosis (ALS). There are numerous observations of astrocyte dysfunction in both humans with ALS and in SOD1(G93A) rodents, a widely studied ALS model. The present study therapeutically targeted astrocyte replacement in this model via transplantation of human Glial-Restricted Progenitors (hGRPs), lineage-restricted progenitors derived from human fetal neural tissue. Our previous findings demonstrated that transplantation of rodent-derived GRPs into cervical spinal cord ventral gray matter (in order to target therapy to diaphragmatic function) resulted in therapeutic efficacy in the SOD1(G93A) rat. Those findings demonstrated the feasibility and efficacy of transplantation-based astrocyte replacement for ALS, and also show that targeted multi-segmental cell delivery to cervical spinal cord is a promising therapeutic strategy, particularly because of its relevance to addressing respiratory compromise associated with ALS. The present study investigated the safety and in vivo survival, distribution, differentiation, and potential efficacy of hGRPs in the SOD1(G93A) mouse. hGRP transplants robustly survived and migrated in both gray and white matter and differentiated into astrocytes in SOD1(G93A) mice spinal cord, despite ongoing disease progression. However, cervical spinal cord transplants did not result in motor neuron protection or any therapeutic benefits on functional outcome measures. This study provides an in vivo characterization of this glial progenitor cell and provides a foundation for understanding their capacity for survival, integration within host tissues, differentiation into glial subtypes, migration, and lack of toxicity or tumor formation.

Advanced technology for storage and transport of purified DNA from umbilical cord blood prior to use in human leukocyte antigens typing and whole-genome microarray analysis.

Two technologies for dry-state, ambient temperature transport of biospecimens were evaluated in this study. Umbilical cord blood (UCB) samples from 4 individuals were transported at ambient temperature using GenPlates, and the DNA recovered was compared with DNA purified directly from granulocytes of the same UCB samples. GenTegra™ DNA tubes were then used to transport the DNA from California to North Carolina and New Zealand, either immediately after drying or following 30 days of storage at 25°C and 76°C. The integrity of the recovered DNA was thoroughly tested using 2 human leukocyte antigens (HLA)-typing techniques (bead array and sequencing), as well as microarray-based whole-genome scanning. HLA-typing results were the same for all samples whether the DNA had been stored for 3 days during transport or 30 days at either 25°C or 76°C. There were no differences in the HLA-typing results of DNA recovered from UCB samples stored in GenPlates compared with DNA extracted directly from granulocytes. Moreover, the microarray analysis revealed call rates of >99.5% for every sample, regardless of storage method, with a statistical concordance of 99.99% between the UCB samples stored in GenPlates compared with DNA extracted directly from granulocytes. These results indicate that both GenPlates and GenTegra are viable methods of storing and transporting UCB (stem cell) biospecimens in a dry state. The quality and quantity of DNA recovered using both technologies are sufficient for complex genotyping using a number of different methods.