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Numerical Methods for the Design and Description of In Vitro Expansion Processes of Human Mesenchymal Stem Cells

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Digital Twins

Part of the book series: Advances in Biochemical Engineering/Biotechnology ((ABE,volume 177))

Abstract

Human mesenchymal stem cells (hMSCs) are a valuable source of cells for clinical applications (e.g., treatment of acute myocardial infarction or inflammatory diseases), especially in the field of regenerative medicine. However, for autologous (patient-specific) and allogeneic (off-the-shelf) hMSC-based therapies, in vitro expansion is necessary prior to the clinical application in order to achieve the required cell numbers. Safe, reproducible, and economic in vitro expansion of hMSCs for autologous and allogeneic therapies can be problematic because the cell material is restricted and the cells are sensitive to environmental changes. It is beneficial to collect detailed information on the hydrodynamic conditions and cell growth behavior in a bioreactor system, in order to develop a so called “Digital Twin” of the cultivation system and expansion process. Numerical methods, such as Computational Fluid Dynamics (CFD) which has become widely used in the biotech industry for studying local characteristics within bioreactors or kinetic growth modelling, provide possible solutions for such tasks.

In this review, we will present the current state-of-the-art for the in vitro expansion of hMSCs. Different numerical tools, including numerical fluid flow simulations and cell growth modelling approaches for hMSCs, will be presented. In addition, a case study demonstrating the applicability of CFD and kinetic growth modelling for the development of an microcarrier-based hMSC process will be shown.

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Abbreviations

CC:

Collagen-coated

CFD:

Computational Fluid Dynamics

DMEM:

Dulbecco’s Modified Eagle Medium

DSP:

Downstream processing

ECM:

Extracellular matrix

bFGF:

Basic fibroblast growth factor

FBS:

Fetal bovine serum

GMP:

Good manufacturing practice

hASC:

Human adipose tissue-derived stromal/stem cells

hBM-MSC:

Human bone marrow-derived mesenchymal stem cells

hMSCs:

Human mesenchymal stem cells

hPL:

Human platelet lysate

HGF:

Hepatocyte growth factor

HSB:

Hemispherical-bottom bioreactor

LDA:

Laser Doppler Anemometry

LES:

Large Eddy Simulations

αMEM:

Modified Eagle Medium

MC:

Microcarrier

MCB:

Master Cell Bank

MRF:

Moving reference frame

OTR:

Oxygen transfer rate

PIV:

Particle Image Velocimetry

PS:

Polystyrene-based

RB:

Round-bottom bioreactor

RMSD:

Root mean square deviation

SIMPLE:

Semi-implicit method for pressure-linked equations

SM:

Sliding mesh

SU:

Single use

UCM:

Umbilical cord-derived mesenchymal stem cells

USP:

Upstream processing

VEGF:

Vascular endothelial growth factor

VOF:

Volume of fluid

WCB:

Working Cell Bank

Amn (mmol/L):

Ammonium concentration

DO2 (m2/s):

Oxygen diffusivity

DR (m):

Vessel diameter

EF :

Expansion factor

F (N):

Force

Glc (mmol/L):

Glucose concentration

h/H L :

Geometrical ratio between a certain height and the liquid height

h R /D R :

Geometrical ratio between impeller installation height and the vessel diameter (= off-bottom clearance)

HL (m):

Liquid height

H L /D :

Geometrical ratio between liquid height and vessel diameter

kat (d-1):

Cell attachment constant

kdet (d-1):

Cell detachment constant

KAmn (mmol/L):

Inhibition constant of ammonium

KGlc (mmol/L):

Monod constant of glucose

KLac (mmol/L):

Inhibition constant of lactate

Lac (mmol/L):

Lactate concentration

N (rpm):

Impeller speed

Ns1u (rpm):

Lower limit of Ns1 suspension criterion

Ns1 (rpm):

1s or just suspended criterion (=Njs)

PDL :

Population doubling level

P/V (W/m3):

Specific (volumetric) power input

pAmn (mmol/cell/d):

Specific ammonium production rate (growth-independent)

pLac (mmol/cell/d):

Specific lactate production rate (growth-independent)

qAmn (mmol/cell/d):

Specific ammonium production rate (growth-dependent)

qGlc (mmol/cell/d):

Specific glucose consumption rate

qLac (mmol/cell/d):

Specific lactate production rate (growth-dependent)

Re :

Reynolds number

r/R :

Dimensionless radial coordinates

tc (s):

Contact time

tcir (s):

Particle circulation times

td (d):

Doubling time of cell population

tl (d):

Lag or cell adaption time

tres (s):

Particle residence time

utip (m/s):

Impeller tip speed

\( \overrightarrow{u} \)(m/s):

Velocity vector in x-direction

Vmin (mL):

Minimal working volume

Vmax (mL):

Maximum working volume

\( \overrightarrow{v\ } \) (m/s):

Velocity vector in y-direction

\( \overrightarrow{w} \)(m/s):

Velocity vector in z-direction

XA (cells/cm2):

Cell concentration on surface

Xmax (cells/cm2):

Maximum cell concentration on surface

XSus (cells/mL):

Cell concentration in suspension

XV (cells/cm2):

Cell concentration of viable cells (XSus + XA)

YLac/Glc (mmol/mmol):

Lactate yield per glucose equivalent

YX/O2 (1/mmol):

Yield coefficient/cells per mmol oxygen

α :

Cell adaption phase coefficient

α MC :

MC volume fraction

δ Glc :

Step response in glucose balance to avoid negative glucose values (δGlc = 0 or 1)

ηL (Pa s):

Dynamic viscosity of the liquid

π :

Mathematical constant (≈ 3.1415)

ρL (kg/m3):

Density of the liquid

τnn (Pa):

Local normal stress

τnt (Pa):

Local shear stress

μ (1/d):

Specific growth rate

μmax (1/d):

Maximum specific growth rate

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  1. Zurich University of Applied Sciences – Institute of Chemistry and Biotechnology, Wädenswil, Switzerland

    Valentin Jossen, Dieter Eibl & Regine Eibl

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    Christoph Herwig

  2. Institute of Bioprocess and Biosystems Engineering, Hamburg University of Technology, Hamburg, Germany

    Ralf Pörtner

  3. Institute of Bioprocess and Biosystems Engineering, Hamburg University of Technology, Hamburg, Germany

    Johannes Möller

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Jossen, V., Eibl, D., Eibl, R. (2020). Numerical Methods for the Design and Description of In Vitro Expansion Processes of Human Mesenchymal Stem Cells. In: Herwig, C., Pörtner, R., Möller, J. (eds) Digital Twins. Advances in Biochemical Engineering/Biotechnology, vol 177. Springer, Cham. https://doi.org/10.1007/10_2020_147

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