Performance evaluation of a novel conceptual bioprocess for clinically-required mass production of hematopoietic cells

Abstract

Objective

The novel engineered bioprocess, which was designed and modeled to provide the clinically relevant cell numbers for different therapies in our previous work (Kaleybar et al. Food Bioprod Process 122:254–268, https://doi.org/10.1016/j.fbp.2020.04.012, 2020), was evaluated by using U937 as hematopoietic model cells.

Results

The culture system showed a 30-fold expansion of U937 cells in one-step during a 10-day culture period. The cell growth profile, the substrate and oxygen consumptions, and byproduct formations were all in agreement with the model predications during 7 days. The cell proliferation decrease after 7 days was attributed to optional oxygen limiting condition in the last days of culture. The bioreactor culture system revealed also a slight enhancement of lactate dehydrogenase (LDH) production as compared to the 2D conventional culture system, indicating the low impact of shear stress on cellular damage in the dynamic system.

Conclusions

The results demonstrated that the conceptual bioprocess for suspended stem cell production has a great potential in practice although additional experiments are required to improve the system.

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Data availability

 All data generated or analyzed during this study are included in this article.

Abbreviations

C :

Molar concentration in solution (pmol/ml or M)

C x :

Cellular density (cells/ml)

D :

The bioreactor diameter (cm)

d m :

Mixer’s diameter (cm)

H :

Height of bioreactor in contact with liquid phase (cm)

H i :

Henry’s constant for solubility of gas i in liquid (Pi/Ci*), (Pa ml/pmol)

K l a :

Mass transfer coefficient (h−1)

n :

Number of repetitions

P :

Pressure (Pa)

pH:

Acidity (–)

\({\overline{q}}\) :

Cell’s specific rate of substrate consumption or metabolite production (pmol/cell day)

t :

Time (h)

V :

Volume (ml)

μ :

Cellular specific growth rate (h−1)

ν :

Stirrer tip speed (π·dm·rpm/60) (m/s)

0:

First value at t = t0

Amm.:

Ammonia

crit.:

Critical

Glu.:

Glucose

initial:

Beginning value at each phase

Lac.:

Lactate

O:

O2

P:

Product or metabolite

S:

Substrate

X:

Cells

*:

Saturated concentration

+:

Positive (for pressure)

max.:

Maximum

DO:

Dissolved oxygen saturation percent in liquid (100Co/Co*), (–)

exp.:

Experimental data

HSC:

Hematopoietic stem cell

LDH:

Lactate dehydrogenase

OTR:

Oxygen transfer rate (Kla)O·(Co* − Co), (pmol/ml h)

rpm:

Stirrer rounds per minute (min−1)

SEM:

Standard error of the mean

STB:

Stirred tank bioreactor

vvm:

Volume of air per unit volume of medium per minute

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Acknowledgements

This work was supported by Iran National Science Foundation (INSF) (No. 96000408). The authors thank all kindly supports.

Funding

This study was partially funded by Iran National Science Foundation (INSF) (No. 96000408). The authors declare that they have no other funding source.

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Correspondence to Ali Baradar Khoshfetrat.

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Kaleybar, L.S., Khoshfetrat, A.B., Rahbarghazi, R. et al. Performance evaluation of a novel conceptual bioprocess for clinically-required mass production of hematopoietic cells. Biotechnol Lett (2021). https://doi.org/10.1007/s10529-020-03062-1

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Keywords

  • Conceptual bioprocess
  • Mass production
  • Sequencing batch aeration
  • Stirred tank bioreactor
  • Suspended stem cell