Attachment II NCC Task 1 SOW FINAL.pdf

TASK ORDER 1 STATEMENT OF WORK

NIEHS DIVISION OF THE NATIONAL TOXICOLOGY PROGRAM

HUMAN NEURAL CREST CELL MIGRATION ASSAY DATA COLLECTION (TASK 1)

1.0. INTRODUCTION: Under this task order, the Contractor shall run the human neural crest cell (NCC) migration assay and collect data on 115 environmental compounds . All services, labor, equipment, and supplies provided under this task order shall be performed under accordance with the terms and conditions of the contract Statement of Work (SOW) and task order SOW.

2.0. TASK: The contractor shall evaluate the neurodevelopmental effects of 115 environmental compounds using two (2) neurodevelopmental endpoints in the NCC: 1) NCC migration; 2) Viability of migrating NCCs.

2.1. Chemicals: All test compounds and dimethylsulfoxide (DMSO) will be supplied by the NIEHS/DNTP blinded in a one-time shipment. All other assay controls shall be supplied by the contractor. The Contractor shall be responsible for supplying all other reagents, equipment and facilities needed to conduct the specific assays.

2.2. Differentiation of hiPSCs to NCC: The contractor shall differentiate hiPSC into migrating NCCs using established differentiation protocols. Morphological characterization of NCC shall be performed using FACS or immunocytochemistry using the NCC marker HNK-1, p75, AP-2 and nestin.

2.3. Migration assay data collection: Neural crest cells shall be seeded into 96-well plates in N2-S medium containing FGF2 and EGF. Cells shall be seeded around stoppers to create a circular cell-free area. Stoppers shall be removed after 24 hours to allow cells to start migration on day of migration 0’ (DoM0’). On DoM1’ cells shall be exposed for 24 hours to test compounds; eight concentrations shall be assayed as well as a DMSO vehicle control. Test concentration shall be prepared in a 1:3 dilution series from a starting concentration of 100 µm. Thereafter, cells shall be stained with calcein-AM and H-33342 and high-content imaging shall be performed (Cellomics ArrayScanVTI; Thermo Fisher Scientific). Total number of viable (calcein-AM and H-33342 double labeled) NCC that have migrated into the region of interest (ROI) shall be determined using high content imaging.

3.0. DATA RIGHTS: See IDIQ base contract.

4.0. DELIVERABLES AND REPORTING REQUIREMENTS: See IDIQ base contract

4.1. Required Reports: A report of completion for Task 1 shall provide a summary of the methods used and the results obtained in sufficient detail and quality to meet the requirements demonstrating completion of the task. The report of completion shall be delivered in both hard and electronic formats (MS Word) for review and comment by the Contracting Officers Representative (COR). The report of completion shall include but not be limited to:

4.1.1. A detailed description of all experimental procedures

4.1.2. A full set of all raw data, summarized/normalized data

4.2. Contract Management Reports: See IDIQ base contract.

4.3. Periodic Status Reports: See IDIQ base contract.

4.4. General Reporting Requirements: See IDIQ base contract.

4.5. Electronic Deliverables: See IDIQ base contract.

5.0. PLACE OF PERFORMANCE: See IDIQ base contract.

6.0. PERIOD OF PERFORMANCE: 12 months.

7.0. Section 508 Requirements for Deliverables: See IDIQ base contract.

8.0. CONTRACTOR FURNISHED MATERIALS: See IDIQ base contract.