Tis Xell Regeneration System Scaffolds Accelerate Your

Tis. Xell Regeneration System & Scaffolds Accelerate Your Cell Proliferation By

Quin. Xell Technologies Ltd • founded in 2011 as a subsidiarity of Quintech Life Sciences • located in Singapore • focus on Regenerative Medicine • Research on Tis. Xell Regeneration System was initiated as a joint project between the National University of Singapore (NUS) and Singapore Polytechnic (SP), with the National University Hospital (NUH) providing samples for applications • Scientific papers supporting the benefits of biaxial bioreactor on functional tissue growth on 3 D scaffolds have been published for more than 10 years

Application Fields • Regenerative medicine • Tissue engineering • Translational medicine • Tissue / organ transplant

Cell Types • Bone Cells • Stem Cells • Primary Cells • Muscle Cells • Heart cells • Bioengineered Cells…

The Novel Approach Type Advantages Tis. Xell Regeneration System Integrated perfusion system allows for exchange between the vessel and reservoir with maximal mass transfer under low sheer force Biaxial revolution of the culture vessel led to more homogenous cellular and ECM deposition – at the surface and the deeper core of the scaffolds Cellular scaffolds were secured by stainless steel needles to avoid scaffold and vessel wall collisions

The Novel Approach Tis. Xell Regeneration System is unique in its bi-directional revolution. The biaxial bioreactor rotates simultaneously in two independent axes, allowing it to spin and tumble simultaneously. If required, the axes can be controlled to function as a single axis system The gyroscopic movement leads to an improved flow of fluids within the bioreactor for accelerated cell proliferation

The Tis. Xell System

Tis. Xell in Motion

Tis. Xell Novel Approach …(1) Innovative Design • Independently controlled drive axes for biaxial, single axis and swing modes • Continuous perfusion with integrated, selectable speed peristaltic pump • Oxygenator unit to facilitate gaseous exchange • 5 -port medium reservoir with sensor bank to house optional temperature, p. H and oxygen probes • Heat isolated system drivers and controller in separate console • System may be placed in a CO 2 incubator or used with a process controller

Tis. Xell Novel Approach …(2) Advantages in Arm Assembly – Weight-balanced rotary arm for 500 ml or 1000 ml spherical chamber – For smaller volume requirements, a 50 ml “pancake” chamber with dual-stacker is available – Built-in sensors to detect arm angle and positions for fully programmable operations – Tube holders to secure excess tubing for obstruction free operation

Tis. Xell Novel Approach …(3) Modular Chamber • • • 50, 500 and 1000 m. L spherical chamber with detachable base to double up as a chamber stand Dual chamber-cap design for easy loading and uploading of tissue scaffolds without removing tubing Oxygenator / Gas membrane for gas diffusion Sterile rotary couplings for controlled continuous perfusion of nutrients through the culture chamber Transparent, unobstructed view of tissue constructs during operation Optional rotary core vessel with increased hydrodynamic shear provides additional stimulus to enhance cell activity

Tis. Xell Novel Approach …(4) Flexible Operation Modes – Biaxial, uniaxial and swing modes can be independently selected – Auto, jog and manual functions can be pre-set to various speeds, directions and arm angles to optimize cell culture conditions – User-friendly remote touch screen controller for complete management of system while unit is inside a laminar flow chamber or a CO 2 incubator

NEW! 50 ml Chamber Small 50 ml vessel for low volume / small scaffold applications Designed for dynamic seeding with “cell concentrator” that concentrate cells around scaffold during seeding loop operations. Dual stacker for simultaneously operating two chambers (in continuous perfusion)

Scaffolds PCL – the ideal material for applications in the Tis. Xell Regeneration System • 3 D inter-woven polycaprolactone • Pores are inter-connected and the polymer has been regarded as cell tissue compatible • Our scaffolds are biocompatible, showing slow degradation and assimilation of the polymer over 24 months • That leads to perfect bone healing, provides adequate mechanical support and maintains the shape of the skull without problems • The scaffolds are durable, load bearing and exhibit slow degradation - ideal for design that requires stress transfer to surrounding cells

Different shapes • available in different sizes and shapes • Specialized shapes so as: - mesh - plug - strip • Customized 3 D shapes feasible

Approvals And of course… …Our scaffolds are FDA Approved for craniofacial applications and certified by the TÜV Rheinland!

BONE APPLICATIONS

Key Benefits for Bone Applications • Homogenous cellular distribution • Rapid stem cell expansion • Promotes new vascularization • High viability • Robust osteogenic differentiation and mineralization • Increased bone formation after transplantation

Rat Femoral Defect Model • Created 7 mm defect • Press-fit 8 mm graft

Healing in TEBG vs. control

CT showing bridging AND mineralization

New blood vessel formation

Mineralization confirmed by histology S S S

Scaling Up • Minipig model • Larger volume, anticipate issues of vascularization • Introduce endothelial progenitor cells (EPC) into cellular mix Mesenchymal stem cells (MSC) Endothelial Progenitor Cells (EPC)

Cells are viable and randomly distributed Tis. Xell supports co-culture of different cell types

Minipig Critical-Size Defect Model q 18 mm segmental defect in tibia q Monitor over 12 months: § X-ray, CT, angiography

CT shows bridging at 3 months f. MSC-EPC MSC 1 mth 3 mth 1 mth

CT-Angiography shows rapid Vascularization f. MSC-EPC MSC 1 mth 3 mth 1 mth

BONE DIFFERENTIATION STUDIES …in vitro

Proliferation: Picogreen assay Tis. Xell: confluent at week 1 Static: far less cells than Tis. Xell group even when they reached confluence at week 4

Cell distribution, viability & shape Tis. Xell High viability throughout 4 wks Homogenous distribution Round shape in wk 4 Static: High viability but necrosis in wk 4 Non-homogenous distribution Spindle shape in wk 4

Differentiation/Mineralization ALP activity and von Kossa staining Tis. Xell group cellular scaffolds have significant higher ALP activity since day 7 and much positive von Kossa staining than static group

Differentiation / mineralization: EM (Week 4)Cellular scaffolds from Tis. Xell are highly mineralized

GENERAL applications

Stem Cell Expansion on Microcarriers …(1) • Tis. Xell has been employed in stem cell research, specifically in cell suspension expansion on microcarriers • Comparison of cell suspension growth in Tis. Xell and spinner flasks are highlighted in the confocal images • Results emphasizes Tis. Xell benefits in accelerating cell growth as reflected by the higher cell number observed as compared to a spinner flask

Stem Cell Expansion on Microcarriers …(2) “A cell seeding density of 5 x 104 cell/ml and we achieved a high cell density of 8 x 105 cell/ml at Day 14 (around 16 fold increase ) by using the Tis. Xell system “ Green dye indicates viable cells

Heart Patch Research …(1) • Heart failure is a major health problem and the leading cause of morbidity and mortality of patients in both industrialized nations and developing world • The regenerative potential of stem cells sparked interest in stem–cell therapy for treating and preventing heart failure have transformed experimental research and led to an explosion in clinical investigation • Researchers at the Nanyang Technological Institute (NTU) grows cells on a customised scaffold to create a "heart patch" for repair of damaged heart tissue

Heart Patch Research …(2) • The diffusion limitation of oxygen transport in the scaled up tissue is overcomed by providing interstitial medium flow to an engineered heart patch using the Tis. Xell system • This approached is expected to enhance tissue assembly by way of improved mass transport. By combining surface laser patterned poly(lactide -co-ε-caprolactone) nanofiber scaffold as substrate and dynamically culturing mesenchymal stem cells in the Tis. Xell system, they improve the viability and proliferation of cells in the scaffolds and achieve minimal cell deterioration.

Implant Research Using Co-Cultures in Animal Studies

Benefits at a glance . . . (1) • Accelerates cell growth, differentiation and cell proliferation, mimicking native extracellular matrix (ECM), hastening time to result • Ideal for slow growing cells and of cell co-cultures • Supports homogenous cell culture at the surface and core of the 3 D scaffolds and microcarriers • Maintains functionality and viability of co-cultures and tissue constructs (implants)

Benefits at a glance . . . (2) • Reduces formation of necrotic neo-tissue that interferes with subsequent implantation process • For stem cells expansion on microcarriers, a 16 -fold cell number increase was in observed on day 14 • In bone cells, Tis. Xell provided a 25 - 35 % increase in cell proliferation • On-going research highlights marked proliferation of cell growth in heart patch research and animal co-culture studies. Tis. Xell undoubtedly, will prove beneficial to a wide ranging cell-based research

Thank you for attention! Helpful links: http: //www. quinxell. com http: //www. rowiak. de Ask for scientific publications!