Stem Cells in A New Era of Cell based Therapies - Creative Biolabs

A stem cell can replicate itself or differentiate into cells that carry out the specific functions of the body. The application of stem cells in regenerative medicine and disease therapeutics is one of the most exciting advances in medical science today. In cell-based therapies, stem cells may play two roles. The first role is as drug-delivery vehicles. The second role is as therapeutic agents themselves. Stem cells also offer opportunities for scientific advances that go far beyond cell-based therapies. Creative Biolabs is dedicated to facilitate the research of stem cells in both basic science and therapeutics development. Please contact us if you are interested in our services or products.

1. Stem Cells in A New Era of Cell-based Therapies www. creative-biolabs.com [email protected]

2. Introduction of Stem Cells What Defines A Stem Cell? Classification and Characteristics of Stem Cells Stem Cells Categorized by Regenerative Applications Application of Stem Cells Applications of Stem Cells in Regenerative Medicine Roles of Stem Cells in Cell-based Therapies Examples of Stem Cell-Based Therapy in Development Applications of Stem Cells in Basic Research Services & Products from Creative Biolabs Stem Cell Services STEMOD™ Disease Cell Models TABLE OF CONTENTS 1 2 3

3. What Defines A Stem Cell? Endless self-renewal , with no limitations or maximum number of replications. Capability to differentiate into any tissue type and cell line in the body. Reference: Understanding Stem Cells: An Overview of the Science and Issues from the National Academies, Washington, DC: National Academies Press, 2016 A stem cell can be defined by two specific characteristics: 3

4. Classification and Characteristics of Stem Cells By Potency Totipotent Stem Cell - can give rise to an entire organism. Pluripotent Stem Cell - can give rise to all tissues. Multipotent Stem Cell - can give rise to discrete cell types. Oligopotent Stem Cell - can differentiate into a few different cell types. Unipotent Stem Cell - can only produce cells of one kind. By Origin Embryonic Stem Cell - pluripotent, risen from the inner cell mass of a pre-implantation embryo. Embryonic Germ Cell - derived from primordial germline cells. Adult Stem Cell – multipotent , found in organs that need a constant supply of cells. By Cell Surface Markers BMPR [MSC] CD4 [WBC] CD8 [WBC] CD34 [HSC, EPC] CD38 [WBC] CD44 [MSC] Lin [HSC, MSC] Sca-1 [HSC, MSC] Stro-1 [HSC, MSC] 4

5. Stem Cells Categorized By Regenerative Applications Embryonic Stem Cell (ESC) Tissue Specific Progenitor Stem Cell (TSPSC) Mesenchymal Stem Cell (MSC) Umbilical Cord Stem Cell (UCSC) Bone Marrow Stem Cell (BMSC) Induced Pluripotent Stem Cell (iPSC) 5

6. Applications of Stem Cells in Regenerative Medicine Modified from Reference: Ranjeet Singh Mahla , International Journal of Cell Biology, 2016 6 ( i ) Improvement of spinal cord injury (ii) Regeneration of retinal sheet (iii) Generation of retinal ganglion cells (iv) Healing of heart defects (v) Hepatic cell formation (vi) Formation of insulin secreting 𝛽 -cells (vii) Cartilage lesion treatment (viii) Regeneration of pacemaker (ix) In vitro gametogenesis 1 ( i ) Treatment of diabetes and retinopathy (ii) Neurodental therapeutic applications (iii) Restoration of cognitive functions (iv) Brain and cancer treatment (v) Ear acoustic function restoration (vi) Regeneration of intestinal mucosa (vii) Treatment of vision defects (viii) Muscle regeneration (ix) Regeneration of fallopian tube 2 ( i ) Regeneration of bladder tissue (ii) Muscle regeneration (iii) Regeneration of teeth tissue (iv) Healing of orthopedic injuries (v) Recovery from muscle injuries (vi) Hear scar repair after attack 3 ( i ) T1DM and T2DM treatment (ii) SLE (autoimmune disease) treatment (iii) Application for HI treatment (iv) Krabbe’s disease treatment (v) Hematopoiesis in neuroblastoma 4 ( i ) Treatment of anemia and blood cancer (ii) Retroviral therapy (iii) Correction of neuronal defects (iv) Generation of functional platelets (v) Alveolar bone regeneration (vi) Regeneration of diaphragm tissue 5 ( i ) Regeneration of kidney tissue (ii) Vision restoration in AMD (iii) Treatment of placental defects (iv) Treatment of brain cortex defects (v) ASD and autism treatment (vi) Treatment of liver and lung disease (vii) Generation of serotonin neurons (viii) Regeneration of pacemaker 6 (1) ESCs, (2) TSPSCs, (3) MSCs, (4) UCSCs, (5)BMSCs, (6) iPSCs Healthy donor Patient

7. Roles of Stem Cells in Cell-based Therapies Stem cells as drug-delivery vehicles Stem cells as therapeutics Activation of antigen-specific immunity Endogenous tissue repair Stem cell-based gene therapy Stem cell-based gene editing Delivery of prodrug -converting enzymes Delivery of apoptosis-inducing agents Delivery of oncolytic viruses Reference: Erin A. Kimbrel and Robert Lanza , Nature reviews, 2020 7

8. Examples of Stem Cell-Based Therapy in Development Reference: Erin A. Kimbrel and Robert Lanza , Nature reviews, 2020 8 Stem Cells as Drug-delivery Vehicles to Treat Cancer Approach Stem cells Engineering feature Cancer Clinical trial ID Enhanced delivery of chemotherapy HB1.F3-CD (myc immortalized fetal NSC line) Cytosine deaminase enzyme to convert 5-FC to 5-FU High-grade glioma NCT01172964 HB1.F3-CD NSC Carboxylesterase to convert irinotecan to potent SN-38 High-grade glioma NCT02192359 Targeted induction of apoptosis Autologous bone marrow MSCs HSV-tk suicide gene Advanced gastrointestinal adenocarcinoma NCT02008539 Umbilical cord MSCs Expression of TRAIL Stage IIIb/IV metastatic lung adenocarcinoma NCT03298763 Delivery of oncolytic virus HB1.F3-CD NSCs CRAd-Survivin-pk7 oncolytic adenovirus Malignant gliomas NCT03072134 Adipose MSCs Thyroidal sodium iodide symporter -expressing oncolytic measles virus Recurrent ovarian cancer NCT02068794 Delivery of checkpoint inhibitor HSCs Conjugated platelets with attached PD-1 antibodies Leukaemia NA (preclinical stage)

9. Examples of Stem Cell-Based Therapy in Development Stem Cells for Induction of Antigen-specific Antitumour Immunity 9 Reference: Erin A. Kimbrel and Robert Lanza , Nature reviews, 2020 S tem cells R eceptor Antigen specificity (+ featu r e) I ndication S tage of d e v elopment HSCs TCR N Y -ESO- 1 (wit h sr39TK suicid e gene) Ovarian , fallopia n tube, peritonea l cancer Phas e I clinica l trial (NCT03691376) KK10 HIV P r eclinica l testin g in humanize d mice HSCs CAR CD 4 (wit h C4 6 fusion) Simian/human immunodeficienc y virus Non-huma n primate testing Adipos e MSCs CAR GD 2 (wit h TRAIL) Glioblastoma I n vit r o testing iPSC-derived T cells CAR CD19 B cel l leukaemia P r eclinica l testin g in xenog r af t model iPSC-derived N K cells CAR Mesothelin Ovaria n cancer P r eclinica l testin g in xenog r af t model iPSC-derived mac r ophages scFv CD20 B cel l leukaemia P r eclinica l testin g in xenog r af t model

10. Examples of Stem Cell-Based Therapy in Development Reference: Erin A. Kimbrel and Robert Lanza , Nature reviews, 2020 Genetically Edited Stem Cells in Clinical Trials And in Preclinical Development 10 Stem cells Locus/loci edited Editing method Purpose or indication S tudy details In preclinical development hESCs β 2 -Microglobulin ( B2M ) AAV Generation of HLA- A , HLA- B and HLA- C null PSCs with overexpressed HLA- E to escape allogeneic immunogenicity Preclinical studies in humanized mice Human iPSCs B2M, CIITA CRISPR–Cas9 Generation of HLA class I and II null PSCs with overexpressed CD47 to escape allogeneic immunogenicity Preclinical studies in humanized mice Human iPSCs HLAA, HLAB, HLAC CRISPR–Cas9 Allele- specific HLA- A , HLA- B and HLA- C disruption to make HLA class I pseudo- homozygous PSCs or disruption of HLA- A and HLA- B with preservation of HLA- C to avoid allogeneic immunogenicity Preclinical studies in immunodeficient mice with human CD8 + T cell challenge Human iPSCs Various AAV, ZFN, TALENs, CRISPR Correction of genetic mutations in patient- specific iPSCs to potentially treat monogenic diseases In vitro proof of concept and Reviewed early preclinical development In clinical development Autologous HSCs BCL11A enhancer region ZFN Re- expression of fetal haemoglobin to treat β- thalassaemia Phase I/II clinical trial (NCT03432364) CRISPR–Cas9 Re- expression of fetal haemoglobin to treat β- thalassaemia Phase I/II clinical trial (NCT03655678) CRISPR–Cas9 Re- expression of fetal haemoglobin to treat sickle cell anaemia Phase I/II clinical trial (NCT03745287) Autologous HSCs CCR5 ZFN Knockout CCR5 to block HIV entry into T cells derived from edited HSCs Phase I/II clinical trial (NCT02500849)

11. Applications of Stem Cells in Basic Research Stem Cells Study Human Development Model Human Diseases Develop New Drugs 11

12. Services from Creative Biolabs Induced Pluripotent Stem Cell Service Mesenchymal Stem Cell Service Organoid and 3D Bioprinting Stem Cell Genome Editing Stem Cell Assay Development Disease-specific Stem Cell Therapy Development iPSC-derived Cell Therapy Development Stem Cell Facilitated Drug Discovery Basic Research Services Therapy Development Services 12

13. STEMOD™ Disease Cell Models 13 Cardiovascular System STEMOD™ LQT1 Disease Model STEMOD™ LQT2 Disease Model STEMOD™ LQT3 Disease Model STEMOD™ LQT8 Disease Model STEMOD™ LAT14 Disease Model STEMOD™ LQT15 Disease Model STEMOD™ CPVT1 Disease Model STEMOD™ CPVT2 Disease Model STEMOD™ DCM Disease Model Nervous System Ocular System STEMOD™ AD Disease Model STEMOD™ ALS Disease Model STEMOD™ DS Disease Model STEMOD™ HD Disease Model STEMOD™ PD Disease Model STEMOD™ PPND Disease Model STEMOD™ RTT Disease Model STEMOD™ SCA Disease Model STEMOD™ SMA Disease Model STEMOD™ Tauopathies Disease Model STEMOD™ POAG Disease Model STEMOD™ AMD Disease Model STEMOD™ Bestrophinopathy Disease Model STEMOD™ RP Disease Model

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