ORGANIZATIONAL HIERARCHY

National Institutes of Health (NIH)

National Eye Institute (NEI)

ROLES / RESPONSIBILITES

Serve as a Lab Technician for a Cell Therapy project in the National Eye Institute (NEI).

Cell Culture (~25%):

Culturing primary human fibroblasts from patients with retinal degenerative diseases.

Catalog and freeze early passages of these fibroblasts.

Reprogram these fibroblasts into induced pluripotent stem cells (iPSCs) using standard available techniques.

Maintain iPSC colonies and freeze their early passages.

Differentiation of patient iPSCs into retinal pigment epithelium (RPE) and endothelial cells and maintenance of those RPE and endothelial cultures. This work will include:

Preparing media, supplements, and reagents needed cell culture work.

Maintaining the cell culture room biosafety level 2 standards and maintaining aseptic conditions in cell culture flow hoods, incubators, and prepare sterile tools needed for work with cell cultures.

The cell culture maintenance work will require working for short periods of times on the weekends and some holidays.

Work to characterize the iPSC colonies for their pluripotency. This will require:

Immunostaining of iPSCs, preparing RNA and cDNA from iPSCs, and performing qRT-PCR assays.

Additionally, will perform three germ layer assay based functional characterization of iPSCs. This work will also include any cell types that act as control for their iPSC work. Other experiments needed to perform these assays utilize one or several of following techniques: immunoblot analysis, immunohistochemistry, immunofluorescence staining, PCR, real time-PCR, RNA and proteins extraction, F ACS, and total tissue resistance measurements.

Perform experiments in the lab involving techniques like immunoblot, qRT-PCR, immunostaining, ELISA, FACS.

Document their results, and compile them for presentations and publications.

Participate in lab meetings and discussions, present their work and the relevant literature, and help in the lab organization.

Help with the other ongoing projects in the lab with the expertise developed on iPSC projects.

Cell Therapy Manufacturing for a CRADA (~10%)

Optimize and develop allo-iPSC-RPE product for comparability studies with auto-iPSC-RPE product

Perform comparability between allo and auto iPSC-RPE using the following assays – flow cytometry, ReShape, SEM, phagocytosis, gene expression, and TER

Manufacture GLP-grade allo-iPSC-RPE for preclinical tox studies

Assist in completing IND-write up for allo-iPSC-RPE

Coordinate tech transfer of allo-iPSC-RPE to the clinical center for use in a phase I clinical trial

GLP/GMP manufacturing of cells (~25%)

Provide conceptual and technical assistance to staff in understanding and implementing the FDA GMP guidelines.

Ensure compliance with GMP; provide guidance to staff.

Lab Management: ordering and supply inventory (~20%)

Oversee lab organization, supplies and reagent inventory, and maintenance of cell culture room.

Oversee repairs, replacement, etc. of laboratory equipment, interacting appropriately with procurement, secretarial, and other staff with related responsibilities.