journal article·2021-01-20Open access

Development of a Biodegradable Microcarrier for the Cultivation of Human Adipose Stem Cells (hASCs) with a Defined Xeno- and Serum-Free Medium

Francesco Muoio, Stefano Panella, Matias Lindner, Valentin Jossen, Yves Harder, Tiziano Moccetti, Regine Eibl, Michele Müller, Tiziano Tallone

Applied Sciences· Volume 11· Issue 3

Abstract

Stirred single-use bioreactors in combination with microcarriers (MCs) have established themselves as a technology that has the potential to meet the demands of current and future cell therapeutic markets. However, most of the published processes have been performed using fetal bovine serum (FBS) containing cell culture medium and non-biocompatible MCs. This approach has two significant drawbacks: firstly, the inevitable potential risks associated with the use of FBS for clinical applications; secondly, non-biocompatible MCs have to be removed from the cell suspension before implantation, requiring a step that causes loss of viable cells and adds further costs and complications. This study aimed to develop a new platform based on a chemically defined xeno- and serum-free cell culture medium and biodegradable MC that can support the growth of human adipose stem cells (hASCs) while still preserving their undifferentiated status. A specific combination of components and manufacturing parameters resulted in a MC prototype, called “BR44”, which delivered the desired functionality. MC BR44 allows the hASCs to stick to its surface and grow in a chemically defined xeno- and serum-free medium (UrSuppe). Although the cells’ expansion rate was not as high as with a commercial non-biodegradable standard MC, those cultured on BR44 maintained a better undifferentiated status in both static and dynamic conditions than those cultured on traditional 2D surfaces.
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Research context

OpenAlex topics: Viral Infectious Diseases and Gene Expression in Insects · 3D Printing in Biomedical Research · Tissue Engineering and Regenerative Medicine

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Foundation for Cardiological Research and Education (FCRE), Cardiocentro Ticino Foundation, 6807 Taverne, Switzerland

Sferalp SA, 6998 Monteggio, Switzerland

Institute of Chemistry and Biotechnology, Competence Center of Biochemical Engineering and Cell Cultivation Technique Zurich University of Applied Sciences, 8820 Wädenswil, Switzerland

Department of Plastic, Reconstructive and Aesthetic Surgery, EOC, 6900 Lugano, Switzerland

Faculty of Biomedical Sciences, Università della Svizzera Italiana, 6900 Lugano, Switzerland

Funding

Swiss Federal Innovation Agency CTI (Commission for Technology and Innovation)

References (71)

Show source-provided references
1. Perspectives for Clinical Translation of Adipose Stromal/Stem Cells10.1155/2019/5858247 ↗
2. Argentati, C., Morena, F., Bazzucchi, M., Armentano, I., Emiliani, C., and Martino, S. (2018). Adipose Stem Cell Translational Applications: From Bench-to-Bedside. Int. J. Mol. Sci., 19.10.3390/ijms19113475 ↗
3. Major clinical application of adipose derived stem cells10.15436/2471-0598.18.1838 ↗
4. Adipose-Derived Stem Cells in Radiotherapy Injury: A New Frontier
5. Skin Tissue Engineering: Application of Adipose-Derived Stem Cells10.1155/2017/9747010 ↗
6. The Use of Adipose-Derived Stem Cells in Selected Skin Diseases (Vitiligo, Alopecia, and Nonhealing Wounds)
7. Manufacturing human mesenchymal stem cells at clinical scale: Process and regulatory challenges10.1007/s00253-018-8912-x ↗
8. A roadmap for cost-of-goods planning to guide economic production of cell therapy products10.1016/j.jcyt.2017.06.009 ↗
9. Current perspectives on the use of ancillary materials for the manufacture of cellular therapies10.1016/j.jcyt.2015.09.010 ↗
10. Serum-free media for the production of human mesenchymal stromal cells: A review10.1111/cpr.12063 ↗
11. Internalized Antigens Must Be Removed to Prepare Hypoimmunogenic Mesenchymal Stem Cells for Cell and Gene Therapy10.1016/j.ymthe.2004.02.012 ↗
12. Composition of Interstitial Fluid10.1093/clinchem/41.10.1522 ↗
13. Interstitial fluid concentrations of glycerol, glucose, and amino acids in human quadricep muscle and adipose tissue. Evidence for significant lipolysis in skeletal muscle10.1172/JCI118043 ↗
14. Adsorption studies of human serum albumin, human gamma-globulins, and human fibrinogen on the surface of p(S/PGL) microspheres10.1163/156856202753419277 ↗
15. Hydrophilic Core-Shell Microspheres: A Suitable Support for Controlled Attachment of Proteins and Biomedical Diagnostics10.1002/mabi.200500138 ↗
16. Affinity separation with polyaldehyde microsphere beads10.1016/S0021-9673(00)91346-3 ↗
17. Development and characterization of Ni-NTA-bearing microspheres10.1002/cyto.10124 ↗
18. Bioerodable PLGA-Based Microparticles for Producing Sustained-Release Drug Formulations and Strategies for Improving Drug Loading10.3389/fphar.2016.00185 ↗
19. Poly(lactic-co-glycolic acid) devices: Production and applications for sustained protein delivery10.1002/wnan.1516 ↗
20. Recent advances in serum-free microcarrier expansion of mesenchymal stromal cells: Parameters to be optimized10.1016/j.bbrc.2015.09.078 ↗
21. Nanostructured injectable cell microcarriers for tissue regeneration10.2217/nnm-2016-0083 ↗
22. Large-scale production of human mesenchymal stem cells for clinical applications10.1002/bab.1006 ↗
23. Biomanufacturing for clinically advanced cell therapies10.1038/s41551-018-0246-6 ↗
24. Biodegradable ECM-coated PCL microcarriers support scalable human early MSC expansion and in vivo bone formation10.1016/j.jcyt.2016.06.016 ↗
25. Gas foamed open porous biodegradable polymeric microspheres10.1016/j.biomaterials.2005.05.081 ↗
26. Alternatives to the use of fetal bovine serum: Serum-free cell culture
27. Fetal bovine serum (FBS): Past-present-future
28. Serum-free media formulations are cell line–specific and require optimization for microcarrier culture10.1016/j.jcyt.2015.05.001 ↗
29. Muñoz, M.S., Confalonieri, D., Walles, H., Van Dongen, E.M.W.M., and Dandekar, G. (2018). Recombinant Collagen I Peptide Microcarriers for Cell Expansion and Their Potential Use as Cell Delivery System in a Bioreactor Model. J. Vis. Exp.
30. Systematic microcarrier screening and agitated culture conditions improves human mesenchymal stem cell yield in bioreactors10.1002/biot.201400862 ↗
31. Improved expansion of human bone marrow-derived mesenchymal stem cells in microcarrier-based suspension culture10.1002/term.1515 ↗
32. Functional Cells Cultured on Microcarriers for Use in Regenerative Medicine Research10.3727/096368910X532792 ↗
33. Scalable microcarrier-based manufacturing of mesenchymal stem/stromal cells10.1016/j.jbiotec.2016.08.007 ↗
34. Influence of Microenvironment on Mesenchymal Stem Cell Therapeutic Potency: From Planar Culture to Microcarriers10.3389/fbioe.2020.00640 ↗
35. Application of human mesenchymal and pluripotent stem cell microcarrier cultures in cellular therapy: Achievements and future direction10.1016/j.biotechadv.2013.03.006 ↗
36. Telomerase Immortalized Human Amnion- and Adipose-Derived Mesenchymal Stem Cells: Maintenance of Differentiation and Immunomodulatory Characteristics10.1089/ten.tea.2008.0205 ↗
37. Jossen, V., Muoio, F., Panella, S., Harder, Y., Tallone, T., and Eibl, R. (2020). An Approach towards a GMP Compliant In-Vitro Expansion of Human Adipose Stem Cells for Autologous Therapies. Bioengineering, 7.10.3390/bioengineering7030077 ↗
38. Fluid Flow and Cell Proliferation of Mesenchymal Adipose-Derived Stem Cells in Small-Scale, Stirred, Single-Use Bioreactors10.1002/cite.201200180 ↗
39. Jossen, V., Eibl, R., Kraume, M., and Eibl, D. (2018). Growth Behavior of Human Adipose Tissue-Derived Stromal/Stem Cells at Small Scale: Numerical and Experimental Investigations. Bioengineering, 5.10.3390/bioengineering5040106 ↗
40. Suspending of solid particles in liquid by agitators10.1016/0009-2509(58)85031-9 ↗
41. Liepe, F., Sperling, R., and Jembere, S. (1998). Rührwerke: Theoretische Grundlagen, Auslegung und Bewertung, Fachhochschule.
42. Scale-up of adipose tissue-derived mesenchymal stem cell production in stirred single-use bioreactors under low-serum conditions10.1002/elsc.201300134 ↗
43. Microcarrier choice and bead-to-bead transfer for human mesenchymal stem cells in serum-containing and chemically defined media10.1016/j.procbio.2017.03.017 ↗
44. Growth, Metabolism, and Growth Inhibitors of Mesenchymal Stem Cells10.1089/ten.tea.2008.0345 ↗
45. Patterns of Amino Acid Metabolism by Proliferating Human Mesenchymal Stem Cells10.1089/ten.tea.2011.0223 ↗
46. Spontaneous decomposition of glutamine in cell culture media10.1016/0014-4827(62)90290-2 ↗
47. The importance of ammonia in mammalian cell culture10.1016/0168-1656(95)00196-4 ↗
48. High-Throughput Screening of Surface Marker Expression on Undifferentiated and Differentiated Human Adipose-Derived Stromal Cells10.1089/ten.tea.2015.0039 ↗
49. Identification and validation of multiple cell surface markers of clinical-grade adipose-derived mesenchymal stromal cells as novel release criteria for good manufacturing practice-compliant production10.1186/s13287-016-0370-8 ↗
50. Review of the adipose derived stem cell secretome10.1016/j.biochi.2013.06.001 ↗
51. Cathepsin S Promotes Human Preadipocyte Differentiation: Possible Involvement of Fibronectin Degradation10.1210/en.2006-0386 ↗
52. The insulin like growth factor and binding protein family: Novel therapeutic targets in obesity & diabetes10.1016/j.molmet.2018.10.008 ↗
53. Insulin-like growth factor I, growth hormone and insulin in white adipose tissue10.1016/j.beem.2005.07.011 ↗
54. Insulin-like growth factor 2 regulates the proliferation and differentiation of rat adipose-derived stromal cells via IGF-1R and IR10.1016/j.jcyt.2018.11.010 ↗
55. Differential effects of IGF-I, IGF-II and insulin in human preadipocytes and adipocytes—Role of insulin and IGF-I receptors10.1016/j.mce.2011.04.005 ↗
56. Control of Adipose Tissue Expandability in Response to High Fat Diet by the Insulin-like Growth Factor-binding Protein-410.1074/jbc.M113.545798 ↗
57. Contributions of the N- and C-terminal domains of IGF binding protein-6 to IGF binding10.1677/jme.1.01547 ↗
58. Key questions and answers about pregnancy-associated plasma protein-A10.1016/j.tem.2012.02.008 ↗
59. Cytokines: Names and Numbers You Should Care About10.2215/CJN.07590714 ↗
60. The Chemokine Superfamily Revisited10.1016/j.immuni.2012.05.008 ↗
61. Pentraxin 3 as a Prognostic Biomarker in Patients with Systemic Inflammation or Infection
62. Plasminogen activator inhibitor-1 modulates adipocyte differentiation
63. PLGA: A unique polymer for drug delivery10.4155/tde.14.91 ↗
64. Gelatin carriers for drug and cell delivery in tissue engineering10.1016/j.jconrel.2014.04.014 ↗
65. Antigenicity and immunogenicity of collagen10.1002/jbm.b.30096 ↗
66. Extracellular matrix-based biomaterial scaffolds and the host response10.1016/j.biomaterials.2016.02.003 ↗
67. Remodelling the extracellular matrix in development and disease10.1038/nrm3904 ↗
68. Recombinant collagen and gelatin for drug delivery10.1016/j.addr.2003.08.008 ↗
69. Theoretical and Practical Issues That Are Relevant When Scaling Up hMSC Microcarrier Production Processes10.1155/2016/4760414 ↗
70. Expansion of human mesenchymal stromal cells on microcarriers: Growth and metabolism10.1002/term.224 ↗
71. Three-dimensional cell culture systems as an in vitro platform for cancer and stem cell modeling10.4252/wjsc.v11.i12.1065 ↗