Module 1: Cell Culture and Cell Phenotyping

The goal of the Cell Culture and Cell Phenotyping Module is to streamline cell culture and phenotyping analyses for vision researchers. To this end, this Module supports the maintenance and authentication of cell lines that are required for rigorous study of a wide range of NEI-focused research activities, including all NEI-funded research projects and new collaborative and pilot projects. The Director and staff of this Module provide training, technical support, and maintenance of key equipment to facilitate scientifically rigorous, cutting edge, vision research.

The Cell Culture arm of this module re-designs a previous cell culture maintenance Facility and transforms it into a high-quality resource that streamlines the workflow for a broad range of vision researchers. It also functions to provide technical expertise to investigators to assist in the establishment of primary cultures and development of new immortalized cell lines. This first arm provides the following essential services:
1) Routine maintenance of immortalized cell lines
2) Maintenance of cryopreserved stocks of cell lines
3) Establishment of primary corneal and retinal cell lines
4) Routine screening for mycoplasma and endotoxin contamination
5) Short tandem repeat DNA profiling for cell line authentication

The Cell Phenotyping arm of this module is designed to provide technical expertise and assistance with ongoing cell phenotyping experiments. This includes overseeing the maintenance of all major pieces of equipment, training of investigators, students, postdocs, and staff on proper use, and guidance on experimental design and data analysis, including:
1) Flow cytometry
2) Cell sorting
3) High throughput cellular analysis
4) Live cell metabolic profiling
5) Microvesicle quantification an size determination

One innovative aspect of this Module is the inclusion of a high throughput imaging cytometer (Celigo), which is not currently available in any other department on campus. A major value of the cytometer lies in the significant reduction in time required for data acquisition and analysis for multiple users. This Module also supports new projects and new applications, such as Dr. Vinod Mootha’s metabolic phenotyping studies of corneal endothelial cells from normal patients and those with Fuchs’ corneal endothelial dystrophy. Similarly, the use of high throughput imaging cytometry wgreatly enhances data collection and slashes the time required for cell phenotyping for many NEI-funded projects. In addition, this module provides the necessary resources for junior faculty and postdocs who are currently preparing their first grant applications to be submitted to NEI.

Collectively, both arms of this Module add scientific value to the work of the individual investigators by reducing the time and effort required for basic cell culture assays such as viability and cell cycle assessments. The incorporation of a flow cytometer and a cell sorter also reduces the cost associated with recurrent use of the main flow cytometry facility on campus, which charges high fees per hour of use and often, makes many of these experiments cost-limited. Moreover, inclusion of a high throughput imaging cytometer allows for many of these same comparisons to be made without the need for trypsinization and extensive cell processing. Inclusion of tunable resistance pulse sensing for the real time analysis of cellular microvesicles adds an additional layer of innovation.