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Building 3D Tissues for Transplantation and Drug Screening

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Until now only the most elite laboratories have been able to build complex high-resolution tissue for drug screening and transplantation. Our technology has made building large 3D tissues as simple as pipetting your favorite cells.

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Building 3D Tissues for Transplantation and Drug Screening

  1. 1. BUILDING 3D TISSUES FOR TRANSPLANTATION AND DRUG SCREENING Consistent, transplantable large-format 3D tissue culture for screening of rare cells in vitro and in vivo. 1
  2. 2. WHY SCREEN THERAPEUTICS IN 3D CELL CULTURE? 2 3D cellular environments give more accurate drug screening results 3D cellular environments induce differential cell gene expression Multi-cell 3D cultures better replicate the in vivo environment Improved human to animal translation
  3. 3. COMMON ROADBLOCKS FOR 3D TISSUE CULTURE ADOPTION 3 Inconsistent Results Inconsistent Spheroid Formation (not all cells form spheroids) Often spontaneously dissociate Spheroids are only a few 1000 cells Limited by Growth Conditions Long cell culture times to establish Hypoxic core limits or alters cell growth High Overhead Specialized training necessary (Masters or PhD) Complex protocols & specialized cell culture methods Limited Use Cases Difficult to effectively transplant spheroids as a tissue Spheroids do not induce spontaneous vascularization Difficult to slice or image for structural studies
  4. 4. VASCULAR SCAFFOLDS MAKE 3D CULTURE SIMPLE 4 Guaranteed 3D growth with any cell type Matrix of your choice may be used Specialized training is not required Standard 96 well plate format Guaranteed 3D cell growth Little to no hypoxia Matrix of your choice may be used Standard culture conditions Specialized training not required 6 month shelf-life 100% engraftment of tumors with only 150,000 cells FACS, IHC, IF and Imaging Compatible Multiple cell types may be layered Standard 96 well plate format Ready to use in 24 hours
  5. 5. UNMATCHED RESOLUTION IN BIOCOMPATIBLE MATERIALS 5 • Prellis Biologics, Inc. uses proprietary laser-based printing methods produce high-fidelity, high-resolution biocompatible 3D structures that match the features of native tissues. • Each vascular feature is hollow, reducing hypoxia, and allowing for cell growth as a large format 3D tissue system. • VascularTissue BlanksTM provide consistent 3D architecture for tissue engineering applications.
  6. 6. SMALL 3D TISSUES WITH HUGE R&D POTENTIAL 6
  7. 7. TISSUE CHIP • >1,000 vascular and microvascular features • Suitable for transplantation • Size: 4 x 4 x 1 mm • Volume: 16 mm3 7
  8. 8. VASCULAR BUNDLE • Suitable for layered and mixed cell seeding • Over 50 vascular branches to support cell niche formation 8
  9. 9. FREEFORM TISSUE STRUCTURES 9 BRIGHTFEILD + HUVECs SUBSTRUCTURE HUVECs stained for Laminin
  10. 10. CURRENT CUSTOMER FEEDBACK 10 "The setup is quick and consistent, and the handling is straightforward.There is a little pain, as in most cases, but a lot less than before. 100% better." -Maria Soloveychik , Ph.D. CEO of SyntheX "The structures are also more consistent, so less variability from that perspective too. Less variability means quicker, with less cost and fewer tests – to results." -Marcus Muench, Ph.D. UCSF Professor "Our cultures took significant time – 10 – 12 days or more, while Prellis is a 24 hour seed cycle to get started with your experiment” – Folarin Erogbogbo, Ph.D. Professor SJSU "You need a constant environment to get a reliable, accurate result across and between experiments.We have to always work in the same reliable substructure.This is a big issue in 3D stem cell field.To do a small screen – variability is a big problem. [Traditional organoids] lack consistency and don't have vasculature." -Jennifer Erwin, Ph.D.Assistant Professor John Hopkins School of Medicine "Their technology provides a radical new dimension in our ability to control cell behaviour by tightly manipulating and controlling micro-scale architecture of 3D culture." - Michael Jones, Ph.D. CEO Cell Guidance Systems
  11. 11. WHAT ARE VASCULAR SCAFFOLDS? 11 Seed cells in seconds Hundreds of laser-printed vascular features Large format 3D tumor growth for up to 4 weeks Oxygen permeable, biocompatible hydrogel that is laser-printed by Prellis Bio technology in designs that mimic vascular-like channels and allow for large format tissue growth under standard conditions. Human IPSCs
  12. 12. ORGANOID GROWTH AT D14 HYPOXIA (PDK1 expression) 12
  13. 13. ORGANOID GROWTH Even with minimal HCT116 cell seeding organoids continue to grow for 3 weeks. Cells remain viable for up to 4 weeks under standard tissue culture conditions. 13
  14. 14. EARLY ANIMAL STUDY DATA 14
  15. 15. HIGH BIOCOMPATIBILITY: SCAFFOLDS TRANSPLANTED WITHOUT CELLS 15 2 WEEKS POST-TRANSPLANTATION 8 WEEKS POST-TRANSPLANTATION RED: CD31 (PNAD) BLUE: DAPI GREEN: PRINTED STRUCTURE RED: CD31 (PNAD) BLUE: DAPI GREEN: PRINTED STRUCTURE Fine capillary growth along and inside printed structure. Larger blood vessel growth hundreds of microns inside the transplanted structure. 20 µm
  16. 16. 16 TRANSPLANTATION OF 150,000 CELL-SEEDED VASCULAR SCAFFOLDS 8 week mouse study – fat pad transplant: Groups: 150,000 tumor cells (A549) + Collagen + vascular blank 150,000 tumor cells (A549) + Matrigel + vascular blank (positive control) 2 million cells per standard Matrigel injection (negative control) Structure only 2 million cells + matrigel 200K cells + collagen 200K cells + matrigel scaffold only 8 week Angiosense imaging
  17. 17. 17 TRANSPLANTATION OF 150,000 CELLS WITH AND WITHOUT SCAFFOLD Ongoing 6 week mouse study scaffold only150,000 tumor cells + Matrigel 150,000 tumor cells + Matrigel + scaffold Week 2 images
  18. 18. 18 TRANSPLANTATION OF 150,000 CELL SEEDED TUMORS 2 million cells + matrigel 200K cells + collagen 200K cells + matrigel scaffold only Representative H&E (8 weeks)
  19. 19. 19 TRANSPLANTATION OF 150,000 CELL SEEDED TUMORS 2 million cells + matrigel 200K cells + collagen 200K cells + matrigel scaffold only Representative IF (8 weeks) RED: CD31 (PNAD) BLUE: DAPI GREEN: PRINTED STRUCTURE
  20. 20. IN VITRO 3D TUMOR STUDIES 20
  21. 21. IN VITRO STUDIES REPLICATE KEY 3D TUMOR INSIGHTS 21 Replicated key study showing that 2D screening can give false positive results, in just 48 hours demonstrating statistical significance with just n = 3.
  22. 22. IN VITRO STUDIES REPLICATE KEY 3D TUMOR INSIGHTS 22 Replicated key study showing that 2D screening can give false negative results, in just 48 hours demonstrating statistical significance with just n = 3.
  23. 23. DOSE RESPONSE CURVE IN TUMOR ORGANOIDS “These are far easier to set up and process than tumor spheroids” – customer feedback from SyntheX 23 Compound A Compound B Dose control control(+++) (++) (+) (+++) (++) (+) Prellis Biologics collaborators independently created and tested A549 tumor organoids. Tumors were grown for 8 days and treated with two peptide-based compounds, demonstrating a dose-dependent response in 3D tissues.
  24. 24. ORGANOID+ HUVEC HUVECs self-organize in 3D on vascular tissue blanks. 24 DAPI STRUCTURE LAMININ DAPI LAMININ LAMININ
  25. 25. VERSATILE 3D TISSUE CULTURE 25 HOWTO BUILDYOUR OWNTISSUE
  26. 26. WWW.PRELLISBIO.COM CONTACT: INFO@PRELLISBIO.COM 26

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