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



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,, 2020), was evaluated by using U937 as hematopoietic model cells.


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.


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.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Data availability

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


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)


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)


First value at t = t0








Beginning value at each phase






Product or metabolite






Saturated concentration


Positive (for pressure)




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


Experimental data


Hematopoietic stem cell


Lactate dehydrogenase


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


Stirrer rounds per minute (min−1)


Standard error of the mean


Stirred tank bioreactor


Volume of air per unit volume of medium per minute


  1. Casas López J, Rodríguez Porcel E, Oller Alberola I, Ballesteros Martin M, Sánchez Pérez J, Fernández Sevilla J, Chisti Y (2006) Simultaneous determination of oxygen consumption rate and volumetric oxygen transfer coefficient in pneumatically agitated bioreactors. Ind Eng Chem Res 45:1167–1171.

    CAS  Article  Google Scholar 

  2. Collins PC, Nielsen LK, Patel SD, Papoutsakis ET, Miller WM (1998) Characterization of hematopoietic cell expansion, oxygen uptake, and glycolysis in a controlled, stirred-tank bioreactor system. Biotechnol Prog 14:466–472.

    CAS  Article  PubMed  Google Scholar 

  3. Deshpande RR, Heinzle E (2004) On-line oxygen uptake rate and culture viability measurement of animal cell culture using microplates with integrated oxygen sensors. Biotechnol Lett 26:763–767.

    CAS  Article  PubMed  Google Scholar 

  4. Jelinek N, Schmidt S, Hilbert U, Thoma S, Biselli M, Wandrey C (2002) Novel bioreactors for the ex vivo cultivation of hematopoietic cells. Eng Life Sci 2:15–18.;2-5

    CAS  Article  Google Scholar 

  5. Kaleybar LS, Khoshfetrat AB, Charoudeh HN (2020) Modeling and performance prediction of a conceptual bioprocess for mass production of suspended stem cells. Food Bioprod Process 122:254–268.

    CAS  Article  Google Scholar 

  6. Khosrowshahi YB, Khoshfetrat AB, Abolghasemi Z, Asenjan KS (2015) Performance evaluation of a proliferation chamber with external stirred conditioning tank for expansion of a suspendable stem cell model. Process Biochem 50:1110–1118.

    CAS  Article  Google Scholar 

  7. Khosrowshahi YB, Khoshfetrat AB, Shamsasenjan K (2016) Ex vivo expansion of hematopoietic stem cells in a proliferation chamber with external stirred conditioning tank: sequential optimization of growth factors. Eng Life Sci 16:254–262.

    CAS  Article  Google Scholar 

  8. Klaus J et al (2007) Effect of CD34+ cell dose on hematopoietic reconstitution and outcome in 508 patients with multiple myeloma undergoing autologous peripheral blood stem cell transplantation. Eur J Haematol 78:21–28.

    Article  PubMed  Google Scholar 

  9. Kresnowati MTAP, Forde G, Chen X (2011) Model-based analysis and optimization of bioreactor for hematopoietic stem cell cultivation. Bioprocess Biosyst Eng 34:81–93.

    CAS  Article  PubMed  Google Scholar 

  10. Kwon J, Kim B-S, Kim M-J, Park H-W (2003) Suspension culture of hematopoietic stem cells in stirred bioreactors. Biotechnol Lett 25:179–182.

    CAS  Article  PubMed  Google Scholar 

  11. Oezemre A, Heinzle E (2001) Measurement of oxygen uptake and carbon dioxide production rates of mammalian cells using membrane mass spectrometry. Cytotechnology 37:153–162.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  12. Ozturk SS, Hu WS (2005) Cell culture technology for pharmaceutical and cell-based therapies. In: Biotechnology and bioprocessing, 1st edn. CRC Press, Boca Raton

  13. Rödling L, Schwedhelm I, Kraus S, Bieback K, Hansmann J, Lee-Thedieck C (2017) 3D models of the hematopoietic stem cell niche under steady-state and active conditions. Sci Rep 7:4625–4625.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  14. Safinia L, Panoskaltsis N, Mantalaris A (2005) Haematopoietic culture systems. In: Chaudhuri J, Al-Rubeai M (eds) Bioreactors for tissue engineering: principles, design and operation. Springer, Dordrecht, pp 309–334

    Google Scholar 

  15. Shayan N, Ebrahimi M, Beiki B, Janzamin E (2012) A non-rotational, computer-controlled suspension bioreactor for expansion of umbilical cord blood mononuclear cells. Biotechnol Lett 34:2125–2131.

    CAS  Article  PubMed  Google Scholar 

  16. Wolfe RP, Ahsan T (2013) Shear stress during early embryonic stem cell differentiation promotes hematopoietic and endothelial phenotypes. Biotechnol Bioeng 110:1231–1242.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

Download references


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


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

Author information



Corresponding author

Correspondence to Ali Baradar Khoshfetrat.

Ethics declarations

Conflict of interest

All the authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

(DOCX 20 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

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).

Download citation


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