D.CSS_Attach1-PD.pdf

Version 11-30-2016-1 FAR Part 11—Describing Agency Needs

PURCHASE DESCRIPTION

1. Statement of Need and Purpose:

This requisition is to acquire expandable lung alveolar epithelial cells for an in-vitro lung tissue model for NCATS ETB SARS-CoV-2 project.

2. Background Information and Objective:

Lung is one of the first organs where respiratory viral infection occurs, and severe diseases have been reported to occur in lung parenchyma. Thus, physiologically relevant in vitro lung models are highly demanded to help finding treatments and studying host responses to virus infections and drug treatments. We have established protocols for barrier tissue models with vascularization using 3D bioprinting of required cellular components in a transwell format. We have been using the unique transwell membrane with the electrospun biodegradable PDLGA scaffolds from Biosurfaces Inc. For the development of in vitro alveoli model, cell proliferation is a limiting factor to secure cell sources. Using electrospun biodegradable Poly(DL-lactide-co-glycolide) (PDLGA) scaffolds in a 96 transwell format allows for better cell-cell communications by a direct contact between alveolar epithelium and vascular endothelium, which improves physiological relevance, and allows for an extensive quantity of drug testing.

The acquisition of expendable lung alveolar epithelial cells (100 X 106 cells per donor; total 3 donors) will provide the program an extensive opportunity for scaled up cell resources from one donor. The objective of this project is to obtain expandable cryopreserved alveolar epithelial stem cells expanded to at least passage 3 with validation data showing appropriate markers of alveolar type1 and type 2 differentiation and ACE-2 expression (Coronavirus receptor) and influenza receptors. It will provide function validation data using transepithelial electrical resistance measurements and a full protocol for generating the cells and models.

3. Generic Name of Product: Epithelial stem cells

4. Purchase Description:

• Brand name: Alveolar epithelial stem cells

• Manufacturer: Biosurfaces

• Catalog number: The item is customized

5. Salient characteristics:

• Cryopreserved alveolar epithelial stem cells expanded to at least passage 3.

• 3 Donors: 100 X 106 cells from each donor.

• Enough cells for at least 200 of 96-well plates from each donor (based on 1 final ~7-fold expansion and seeding 30K cells/well)

• Cells are expected to maintain plasticity to allow differentiation into Alveolar Type 2

(AT2) and Type 1 (AT1) cells.

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Version 11-30-2016-1 FAR Part 11—Describing Agency Needs

• Cells will be characterized for appropriate markers of AT1 and AT2 differentiation and ACE-2 expression (Coronavirus receptor) and influenza receptors.

• Cells demonstrates human air/blood barrier model (alveolar epithelium and endothelium) on 96-well Transwell plates.

• Full protocols will be transferred to NIH. Protocols and any necessary hands-on training will be provided to allow future use directly by NIH.

6. Quantity: 3 Donors: 100 X 106 cells from each donor.

7. Delivery Date: One-time delivery within 4-5 months ARO.

8. Period of Performance: N/A

9. Option(s): N/A

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