1. NON-PROPRIETARY July 2012 11 NASA Ames Research Center ARC ISS Utilization Office Cell Bio Tech Demo Payload Overview POIWG MSFC July 25, 2012

2. NON-PROPRIETARY July 2012 22 Cell Bio Tech Demo Project ManagerNicole Raylnicole.a.rayl@nasa.gov (NASA) Project EngineerLance Ellingsonlance.q.ellingson@nasa.gov (Lockheed Martin) Project Scientist/OpsKevin Sato, Ph.D.kevin.y.sato@nasa.gov (Lockheed Martin) SafetySusan Suffelsusan.suffel@nasa.gov (NASA) Payload Integration ManagerAmy Haasamy.s.haas@nasa.gov (Boeing) Systems Engineer (Lockheed Martin)Linda Timucin Verification Engineer (Lockheed Martin)Robert Phillips Operations/Crew Training (Teledyne Brown)Tony Cox SS&MA (NASA)Robert Burney Test Engineer (NASA)Ricardo Oliveras Payload Test Scientist (Lockheed Martin)Natalya Dvorochkin PARC (Teledyne Brown)Robert Powell Outreach (NASA)Barbara Navarro Schedules (Lockheed MartinRobert Pietrowicz Risk Management (Lockheed Martin)Diana Ly

3. NON-PROPRIETARY July 2012 3 NASA ARC, ISS Utilization Office Cell Bio Tech Demo Background Bioculture System for ISS NASA ARC ISS Utilization Office is developing the next generation incubator that will be dedicated to life science space flight research: ­Cell Culture Module (20 Shuttle missions) heritage with new capabilities ­Mammalian and non-mammalian cell cultures (adherent, suspension, 3D) ­Microbiology cultures (prokaryotic and eukaryotic) Capabilities ­Multiple independent experiment sub-housings ­Closed loop flow path system allowing medium oxygenation and profusion ­Independent temperature control ­3 levels of containment (closed loop plus 2 additional covers) ­Automated systems for injection of fixatives, factors, etc. ­Automated environmental gas supply (not independent) ­Crew access for on-orbit experiment initiation, change-out of specimens and media bags, specimen injection into biospecimen chambers, sub-culturing, sampling, factor injection, and use of other ISS science facilities

4. NON-PROPRIETARY July 2012 44 Experiment Summary Cell Bio Tech Demo will conduct on-orbit tests and demonstrate selected new capabilities to verify their design and function for use in the Bioculture System 1.Demonstrate that the tools and procedures used to introduce or remove specimens from a medium bag can be completed on-orbit without the introduction of a microbial contaminant into the liquid medium 2.Demonstrate the on-orbit functional compatibility of Bioculture Systems components and tools with ISS Wetlab kit items 3.Demonstrate the introduction of a biological specimen into the medium bag and sealing of flow paths using candidate tools 4.Demonstrate use of a candidate tubing cutter and crimp tubing for sealing and maintaining sterility on- orbit 5.Obtain crew feedback concerning components and their on-orbit operation, utilize that feedback in the development of crew procedures. 6.Support NASA education/outreach by distributing the tech demo biological samples obtained as a result on-orbit operations, if possible Cell Bio Tech Demo

5. NON-PROPRIETARY July 2012 55 Payload Flight Summary Spx-2 – Launch and Return  Samples/Specimens –Microbiology Medium (TSB; Media Trays 1 and 2) –Fluorescence-tagged Beads (OptiCell™ Interface Tray) –S. cerevisiae (yeast) in YPD Hardware Configuration Launch: L-4 weeks turnover CTB 1; L-48 hrs turnover CTB 2 –Unpowered, ambient Dragon conditions On-Orbit: Both bags stowed at ISS ambient Facility Resource: MELFI +4 o C Dewar for yeast culture post incubation Return: CTB 1 and CTB 2 at ambient Bags containing yeast cultures in +4 oC Double Cold Bag Turnover bags and specimens to the PD at the Southern California payload turnover location Cell Bio Tech Demo Media Tray 1 Kit (3) Media Tray 2 Kit (3) OptiCell Interface Tray Kit (1) Tubing Cutter Specimen Transfer Tray Kit (3) Tubing Cutter Cell Bio Tech Demo CTB 1 Cell Bio Tech Demo CTB 2

6. NON-PROPRIETARY July 2012 6 Cell Bio Tech Demo On-Orbit Operations Summary

7. NON-PROPRIETARY July 2012 7 Cell Bio Tech Demo Media Bag 1 Media Bag 2 Syringe Invision Connector Crimp Tubing Media Bag Crimp Tubing Nutrient Transfer Tool Media Transfer Container Velcro Tab Media Transfer Tray 1 (3 ea): Ops: Transfer 5 ml of Tryptic Soy Broth (TSB) from Media Transfer Container to Media Bag containing TSB using the syringe-based Sample Tool to simulate starting a cell culture Demo: Maintenance of medium sterility using crimp tool to close tubing Media Transfer Tray 2 (3 ea): Ops: Transfer 10 ml of TSB between Media Bags pre-filled with TSB to simulate specimen transfer, such as subculturing Demo: Maintenance of medium sterility

8. NON-PROPRIETARY July 2012 8 Cell Bio Tech Demo Bead Bag OptiCell™ Sampling Tool OptiCell™ YPD Bag Crimp Tubing Yeast Sample Tool YeastTransfer Container Velcro Tab OptiCell™ Interface Tray (1 Tray, 3 OptiCell™s) : Ops: Transfer fluorescent beads (in phosphate buffered saline (PBS) into an Opticell™ filled with PBS Demo: Use of the Sample Tool for transferring a specimen to a carrier that can be moved to another ISS analysis facility Specimen Transfer Tray (3 ea): Ops: Inoculate a live S. cerevisiae (yeast in stasis in saline) to a medium bag pre- filled with YPD medium using the Sample Tool Demo: Use of Sample Tool for transferring a live yeast to initiate culture growth; Education Outreach element

9. NON-PROPRIETARY July 2012 9 Cell Bio Tech Demo Crew Training: On-board training video Crew Operation: Approximately 2 Hrs –Tray operations (as previously described) –Specimen Transfer Tray ◆ Transfer of yeast-filled YPD bags to MELFI +4 o C Dewar post incubation ◆ Transfer of chilled yeast-filled YPD bags to +4 o C conditioned DCB for return Crew Sessions –Session 1: Media Transfer Trays 1 and 2 ◆ Conduct no later than R-2 weeks, may be conducted earlier –Session 2: OptiCell™ Interface Tray ◆ Conduct no later that R-2 weeks, may be conducted earlier –Session 3: Specimen Transfer Tray ◆ Conduct no earlier than R-2 weeks

10. NON-PROPRIETARY July 2012 10 Cell Bio Tech Demo Specimen Transfer Tray Crew Operations Detailed After the yeast are transferred to the YPD Bag, the bag is removed from the tray and stowed in a separate Ziplock bag; all yeast-filled YPD Bags are stowed in the same Ziplock bag. The Specimen Transfer Kits and the Ziplock bag containing with the yeast bags are returned to the CTB Yeast are incubated for 12 hrs to 24 hrs (tests in progress) At the end of incubation, the Ziplock bag containing the yeast bags is transferred to a MELFI +4 o C Dewar The Ziplock bag containing the yeast bags is transferred to a +4 o C conditioned DCB for return in Dragon