The Robotic AUTOSTEM Platform For GMP-Compliant Manufacture of Bone

Marrow-Derived Mesenchymal Stromal Cells

Mary Murphy

Professor in Regenerative MedicineREMEDI, National University of Ireland Galway

Oct 9, 2019

AutostemDevelopment of closed, scalable and

AUTOmated robotic systems for therapeutic STEM cell manufacturing: aseptic systems

from “Donor-to-Patient”

Mesenchymal Stem/Stromal Cells (MSCs) from Human Bone Marrow

AUTOSTEM Vision

This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement no 667932. 5

AUTOSTEM Project• Development of an automated “factory”

• Large-scale production of therapeutic mesenchymal stem/stromal cells (MSCs)

• GMP- and GAMP- compliant• Cost-effective production of adherent cells

• Produced as a sterile, cryopreserved product in vials

• Allogeneic use

• Modular platform• Adaptable for production of autologous MSCs

• Production of MSC-derived Exosomes/EVs

AUTOSTEM Concept

Autostem pipeline – requirements

• ClosedProcess is performed enclosed in a sterile cabinet

• Robot assistedNo direct interaction between user and product

• Integrative and modularImplementation of automated devices and modules for different steps in the protocol

• High capacityBioreactor based expansion of cells in the multi-liter scale

• Sterile and safeSystematic development of hardware according to GMP and VDI 2221 guidelines

• Supporting the regulatory processes Centralized control software for full platform control and data accessibility

• Engaged with HPRA

AutoStem Cell Harvesting System(Crospon)

7

Platform Concept

8

Automated facility functionality

Open Cell Handling (Critical Step)

Marrow aspiration

Seeding (Bioreactor)

Expanding (Bioreactor)

Freezing

Cell delivery

Sampling Quality control

Harvesting

Filling

© WZL/Fraunhofer IPT

Open cell processing

Autostem pipeline – Process workflow

Bone marrow aspiration

Isolation in Bioreactor

Expansion in Bioreactor

Sampling

Sample Analysis

FillingHarvesting Formulation Storage (-80°C)

Transition

Grade D

Grade A

Autostem pipeline – GMP compliance - HPRA

Platform Representation

Autostem pipeline

Storages for disposables

Filling station

Centrifuge

Central robot arm

Decapper

Vial Hatch

Storage for disposables

Cell counter

Automated sampling

Seeder and expander bioreactors

-80 °C Freezer

Central robot arm

This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement no 667932.

Autostem pipeline – concept and flow

Transition

Grade D Grade A

Bone marrow aspiration Isolation Expansion

Sampling

Sample Analysis

Harvesting

Filling

Formulation

Cryo-preservation

Pick-up and final QC

MANUAL (Stir Tank) vs Automated MSC Production

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1.00

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Cel

l co

nce

ntr

atio

n [

10

^5

cel

ls /

ml]

Time in culture [days]

Manual Automated

MANUAL AUTOMATEDFold increase

20.68 ±0.92 21.49 ±2.56

Population doublings

4.37 ±0.06 4.42 ±0.17

Specific growth rate (h-1)

0.0115 ±0.0002

0.0116 ±0.0005

Doubling time (h)

60.4 ±0.9 59.8 ±2.3

D3 D7 D10

Instrumentation enclosure

Probe

Gen1 Gen2

Autostem Sensors / Tyndall-UCC

Probe in Mobius bioreactor

Sensor Performance

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(a) Lactate sensor in response to lactate titration in 10mM

PBS; (b) lactate sensor calibration plot

(a) Glucose sensor response to glucose

titration; (b) the resulting calibration plot from

these readings

(a) Ammonia sensor CV response to ammonia titration

in 10mM PBS; (b) ammonia sensor calibration plot.

Other aspects

• Biological validation

• MSCs, MSC-derived EVs validated• Fracture/bone repair indications

• Regulatory compliance

• GMP and GAMP documentation

• Cost of Goods Analysis

Autostem: the future

• Establish an AUTOSTEM Pilot Plant at a clinical or central production centre

• Ensure full GMP and GAMP compliance

• Extend functionality to other tissue sources

• Validate the production process with donor tissue

• Regulatory approval

• Clinical testing for equivalence

• Commercialisation

AutoCRAT: the future EU Funded / Under negotiation

“Automated Cellular Robot-Assisted Technologies for translation of discovery-led stem cell therapy development for Osteoarthritis”

• Expansion to production of iPSC-derived MSCs and chondrocytes

• Automated production of extracellular vesicles (EVs)

• Finalise GMP and GAMP compliance of the expanded factory

• Test clinical product for osteoarthritis in horses

AutoCRAT Factory Concept

Autostem Partners

This project has received funding from the European Union’s Horizon 2020 research and

innovation programme under grant agreement no 667932.

Acknowledgements

STARSTEM

We acknowledge funding/scientific input from Abbvie

Working Autostem Factory

https://youtu.be/GMQTK8JpcQ8