An Approach towards a GMP Compliant In-Vitro Expansion of Human Adipose Stem Cells for Autologous Therapies
Human Adipose Tissue Stem Cells (hASCs) are a valuable source of cells for clinical applications (e.g., treatment of acute myocardial infarction and inflammatory diseases), especially in the field of regenerative medicine. However, for autologous (patient-specific) and allogeneic (off-the-shelf) hAS...
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MDPI AG
2020-07-01
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| Series: | Bioengineering |
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| Online Access: | https://www.mdpi.com/2306-5354/7/3/77 |
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| author | Valentin Jossen Francesco Muoio Stefano Panella Yves Harder Tiziano Tallone Regine Eibl |
| author_facet | Valentin Jossen Francesco Muoio Stefano Panella Yves Harder Tiziano Tallone Regine Eibl |
| author_sort | Valentin Jossen |
| collection | DOAJ |
| description | Human Adipose Tissue Stem Cells (hASCs) are a valuable source of cells for clinical applications (e.g., treatment of acute myocardial infarction and inflammatory diseases), especially in the field of regenerative medicine. However, for autologous (patient-specific) and allogeneic (off-the-shelf) hASC-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 hASCs for autologous therapies is more problematic because the cell material changes for each treatment. Moreover, cell material is normally isolated from non-healthy or older patients, which further complicates successful in-vitro expansion. Hence, the goal of this study was to perform cell expansion studies with hASCs isolated from two different patients/donors (i.e., different ages and health statuses) under xeno- and serum-free conditions in static, planar (2D) and dynamically mixed (3D) cultivation systems. Our primary aim was I) to compare donor variability under in-vitro conditions and II) to develop and establish an unstructured, segregated growth model as a proof-of-concept study. Maximum cell densities of between 0.49 and 0.65 × 10<sup>5</sup> hASCs/cm<sup>2</sup> were achieved for both donors in 2D and 3D cultivation systems. Cell growth under static and dynamically mixed conditions was comparable, which demonstrated that hydrodynamic stresses (<i>P/V =</i> 0.63 W/m<sup>3</sup>, <i>τ<sub>nt</sub></i> = 4.96 × 10<sup>−3</sup> Pa) acting at <i>N<sub>s1u</sub></i> (49 rpm for 10 g/L) did not negatively affect cell growth, even under serum-free conditions. However, donor-dependent differences in the cell size were found, which resulted in significantly different maximum cell densities for each of the two donors. In both cases, stemness was well maintained under static 2D and dynamic 3D conditions, as long as the cells were not hyperconfluent. The optimal point for cell harvesting was identified as between cell densities of 0.41 and 0.56 × 10<sup>5</sup> hASCs/cm<sup>2</sup> (end of exponential growth phase). The growth model delivered reliable predictions for cell growth, substrate consumption and metabolite production in both types of cultivation systems. Therefore, the model can be used as a basis for future investigations in order to develop a robust MC-based hASC production process for autologous therapies. |
| first_indexed | 2024-03-10T18:21:49Z |
| format | Article |
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| language | English |
| last_indexed | 2024-03-10T18:21:49Z |
| publishDate | 2020-07-01 |
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| spelling | doaj.art-66e698368cbd4dbc952760e5376981362023-11-20T07:17:58ZengMDPI AGBioengineering2306-53542020-07-01737710.3390/bioengineering7030077An Approach towards a GMP Compliant In-Vitro Expansion of Human Adipose Stem Cells for Autologous TherapiesValentin Jossen0Francesco Muoio1Stefano Panella2Yves Harder3Tiziano Tallone4Regine Eibl5Institute of Chemistry and Biotechnology, Zurich University of Applied Sciences, 8820 Wädenswil, SwitzerlandFoundation for Cardiological Research and Education (FCRE), Cardiocentro Ticino Foundation, 6807 Taverne, SwitzerlandFoundation for Cardiological Research and Education (FCRE), Cardiocentro Ticino Foundation, 6807 Taverne, SwitzerlandDepartment of Plastic, Reconstructive and Aesthetic Surgery, Ente Ospedaliero Cantonale (EOC), 6900 Lugano, SwitzerlandFoundation for Cardiological Research and Education (FCRE), Cardiocentro Ticino Foundation, 6807 Taverne, SwitzerlandInstitute of Chemistry and Biotechnology, Zurich University of Applied Sciences, 8820 Wädenswil, SwitzerlandHuman Adipose Tissue Stem Cells (hASCs) are a valuable source of cells for clinical applications (e.g., treatment of acute myocardial infarction and inflammatory diseases), especially in the field of regenerative medicine. However, for autologous (patient-specific) and allogeneic (off-the-shelf) hASC-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 hASCs for autologous therapies is more problematic because the cell material changes for each treatment. Moreover, cell material is normally isolated from non-healthy or older patients, which further complicates successful in-vitro expansion. Hence, the goal of this study was to perform cell expansion studies with hASCs isolated from two different patients/donors (i.e., different ages and health statuses) under xeno- and serum-free conditions in static, planar (2D) and dynamically mixed (3D) cultivation systems. Our primary aim was I) to compare donor variability under in-vitro conditions and II) to develop and establish an unstructured, segregated growth model as a proof-of-concept study. Maximum cell densities of between 0.49 and 0.65 × 10<sup>5</sup> hASCs/cm<sup>2</sup> were achieved for both donors in 2D and 3D cultivation systems. Cell growth under static and dynamically mixed conditions was comparable, which demonstrated that hydrodynamic stresses (<i>P/V =</i> 0.63 W/m<sup>3</sup>, <i>τ<sub>nt</sub></i> = 4.96 × 10<sup>−3</sup> Pa) acting at <i>N<sub>s1u</sub></i> (49 rpm for 10 g/L) did not negatively affect cell growth, even under serum-free conditions. However, donor-dependent differences in the cell size were found, which resulted in significantly different maximum cell densities for each of the two donors. In both cases, stemness was well maintained under static 2D and dynamic 3D conditions, as long as the cells were not hyperconfluent. The optimal point for cell harvesting was identified as between cell densities of 0.41 and 0.56 × 10<sup>5</sup> hASCs/cm<sup>2</sup> (end of exponential growth phase). The growth model delivered reliable predictions for cell growth, substrate consumption and metabolite production in both types of cultivation systems. Therefore, the model can be used as a basis for future investigations in order to develop a robust MC-based hASC production process for autologous therapies.https://www.mdpi.com/2306-5354/7/3/77human adipose stem cells (hASCs)serum- and xeno-free conditionsUrSuppe stem cell culture mediumautologous therapykinetic growth modelingsegregated and unstructured growth model |
| spellingShingle | Valentin Jossen Francesco Muoio Stefano Panella Yves Harder Tiziano Tallone Regine Eibl An Approach towards a GMP Compliant In-Vitro Expansion of Human Adipose Stem Cells for Autologous Therapies Bioengineering human adipose stem cells (hASCs) serum- and xeno-free conditions UrSuppe stem cell culture medium autologous therapy kinetic growth modeling segregated and unstructured growth model |
| title | An Approach towards a GMP Compliant In-Vitro Expansion of Human Adipose Stem Cells for Autologous Therapies |
| title_full | An Approach towards a GMP Compliant In-Vitro Expansion of Human Adipose Stem Cells for Autologous Therapies |
| title_fullStr | An Approach towards a GMP Compliant In-Vitro Expansion of Human Adipose Stem Cells for Autologous Therapies |
| title_full_unstemmed | An Approach towards a GMP Compliant In-Vitro Expansion of Human Adipose Stem Cells for Autologous Therapies |
| title_short | An Approach towards a GMP Compliant In-Vitro Expansion of Human Adipose Stem Cells for Autologous Therapies |
| title_sort | approach towards a gmp compliant in vitro expansion of human adipose stem cells for autologous therapies |
| topic | human adipose stem cells (hASCs) serum- and xeno-free conditions UrSuppe stem cell culture medium autologous therapy kinetic growth modeling segregated and unstructured growth model |
| url | https://www.mdpi.com/2306-5354/7/3/77 |
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