Cord derived MSC-Like Placenta derived Membranes- Matrix- Cells- MSC-like,

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1 Bank Public/Pvt Bank other cells Ship and distribute Collect and process other samples Support development of novel uses One cord Blood Unit Non stem cells RBC s Plateletes T B Monocytes NK cells Stem & Progenitor cells CD34+ EBC (< 1x10^6) MSC (<1x10^8) others Plasma EBV transformed lines Immortalized progenitors ipsc Cord derived MSC-Like Placenta derived Membranes- Matrix- Cells- MSC-like, Commercial products available

2 ipsc from cord blood Fresh Harvest NYCBB At time of use of unit Confirm consent Data Anonymization Issues HLA typing data From designed bag or tubing Shipping Tracking Identity CD34+ cells or mononuclear fraction Receive at NYSCF ipsc generation 50,000 cells Sendai Electroporation Vectors IP issues 96 well format Media and timeline ipsc testing Pluripotency Differentiation Karyotype Ship and store

3 ipsc One to One One to Many Many to Many Therapy models Autologous Yes No No Donor Self Healthy HLA matched No of lines 1-3 clones 5-10 lines >100lines Regulated as BLA Multiple products Fewer tests and tracking issues Longer time to therapeutic product May be GLP HE exemption Consent straightforward Immune suppression Cost Yes Yes Yes Yes Yes Yes Yes More Even More Yes Possibly Possible Much shorter Unlikely Shorter Unlikely Yes Burdensome Burdensome Unnecessary High/patient Unlikely High per patient Lowest Unlikely Intermediate Intermediate

4 Pros Is a hematologic source a better choice? 1: No of cells is very large so save time in the initial culture period 2: Stem cell populations may be easier material to make ipsc from 3: Cord blood cells less exposed to environmental damage 4: Reported efficiencies higher 5: Electroporation works well 6: Large scale flow based technologies for electroporation available (Maxcyte) 7: Cord and blood banks have optimized collection and storage procedures 8: Collection itself is relatively painless 9: Large no s of ethnically diverse HLA typed samples available today Cons 1: Private cord blood banks do not type and store in single use bags 2: Lentivirus and other transduction technologies not effective in stem cell populations 3: May be biased to mesodermal lineages 4: Some zerofootprint technologies may not work (? Sendai) 5: Propagation and amplification of HSC is difficult

5 Possible Path forward With private cord blood Bank 1: Define a marketing model and revenue stream 2: Confirm consent and plan for HLA typing 3: Identify exact part of sample that will be used 4: Identify a ipsc generation service provider 5: Determine a zerofootprint technology one can use without IP issues 6: Develop an automated scalable process 7: Test storage and shipping issues 8: Define release and quality criteria of pluripotent cells With public cord blood Bank or blood bank 1: Define a agreement and ensure compliance with HIPAA rules 2: Identify exact part of sample that will be used 3: Identify a ipsc generation service provider 4: Determine a zerofootprint technology one can use without IP issues 5: Develop an automated scalable process 6: Test storage and shipping issues 7: Define release and quality criteria of pluripotent cells

6 Why should private cord blood banks consider this 1: Need to find use for cord blood to maintain business model 2: Need additional revenue stream 3: If Proof of principle exists for frozen cord blood samples then the value of stored units gets significantly greater 4: Partnering with an ipsc generation service provider provides route for expansion 5: Offers an opportunity to silence critics of the industry

7 Table 1. Methods for reprogramming human cord blood and/or peripheral blood cells Method Molecule Disposition Validated human blood rep.? Blood rep. efficiency Retroviral DNA Integrated Yes % Lentiviral DNA Integrated Yes % Lentiviral/cre-lox DNA Excisable Not yet N/A Adenovirus DNA Degraded Not yet N/A piggybactransposon DNA Excisable Not yet N/A Episomal vectors DNA Kicked out Yes % Minicircles DNA Yes Not yet N/A Snythetic nucleotides Synthetic mrna Degraded Not yet N/A Synthetic nucleotides Synthetic mirna Degraded Not yet N/A Sendai virus RNA Kicked out Yes % Recombinant protein Protein Degraded Not yet N/A

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9 Strategies for Cord Blood reprogramming Ficoll Gradient Plasma Mononuclear Cells Ficoll-Paque Media Granulocytes & Erythrocytes HSC Sendai Virus Rep. Results UCB-MNC ipsc-p0, TRA-1- ipsc-p1, TRA-1-60 ipsc-p2 60 Nanog/TRA-1-81/DAPI Oct4/TRA-1-60/DAPI Sox2/SSEA-4/DAPI EB

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12 Summary. $$ UCB UCB Donate Private UCB Bank Public UCB Bank $$ Storage Clinic Un-related Transplant Institute Basic Research Biotech. Cell Product Family Related Transplant Personalized Medicine Cord blood bank business model. In private banks, parents pay the bank to store the cord blood. The family owns the cord blood and can decide how it is used. In Public banks, banks pay for the collection, testing, storage and they own the cord blood. The cord blood could be used by anyone who may need it for un-related transplant. The cord blood could also be used for research or for commercially production of CB-derived cells (e.g. mesenchymal stem cell, endothelial progenitor cells)

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