Scalable Culture Strategies for the Expansion of Patient-Derived Cancer Stem Cell Lines
Cancer stem cells (CSCs) have recently raised great interest as a promising biological system for designing effective cancer therapies. The scarcity of CSCs in vivo and the consequent low numbers obtained from biopsies represent a major hurdle to the development of such strategies. It is therefore n...
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| Format: | Article |
| Language: | English |
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Wiley
2019-01-01
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| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2019/8347595 |
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| author | Ana Teresa Serra Margarida Serra Ana Carina Silva Tamara Brckalo Anita Seshire Catarina Brito Michael Wolf Paula M. Alves |
| author_facet | Ana Teresa Serra Margarida Serra Ana Carina Silva Tamara Brckalo Anita Seshire Catarina Brito Michael Wolf Paula M. Alves |
| author_sort | Ana Teresa Serra |
| collection | DOAJ |
| description | Cancer stem cells (CSCs) have recently raised great interest as a promising biological system for designing effective cancer therapies. The scarcity of CSCs in vivo and the consequent low numbers obtained from biopsies represent a major hurdle to the development of such strategies. It is therefore necessary to design robust scalable methods to enable efficient expansion of bona fide CSCs in vitro. Here, we evaluated the applicability of computer-controlled bioreactors combined with 3D aggregate culture and microcarrier technology, widely used in stem cell bioprocessing, for the expansion and enrichment of CSCs isolated from different types of solid tumors—colorectal cancer (CRC) and non-small-cell lung cancer (NSCLC) from two patients. Results show that these culture strategies improved cell expansion and CSC enrichment. Both patient-derived CSC lines were able to grow on microcarriers, the best results being achieved for PPlus 102-L, Pro-F 102-L, Fact 102-L, and CGEN 102-L beads (5-fold and 40-fold increase in total cell concentration for CRC and NSCLC cells, respectively, in 6 days). As for 3D aggregate culture strategy, the cell proliferation profile was donor dependent. NSCLC cells were the only cells able to form aggregates and proliferate, and the flat-bottom bioreactor vessel equipped with a trapezoid-shaped paddle impeller was the most efficient configuration for cell growth (21-fold increase in cell concentration achieved in 8 days). Serum-free medium promotes CSC enrichment in both 3D aggregate and microcarrier cultures. The protocols developed herein for CSC expansion have the potential to be transferred to clinical and industrial settings, providing key insights to guide bioprocess design towards the production of enriched CSC cultures in higher quantity and improved quality. |
| format | Article |
| id | doaj-art-31d27fd4da3047f29bca769bc4c1a8d7 |
| institution | OA Journals |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2019-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-31d27fd4da3047f29bca769bc4c1a8d72025-08-20T02:07:40ZengWileyStem Cells International1687-966X1687-96782019-01-01201910.1155/2019/83475958347595Scalable Culture Strategies for the Expansion of Patient-Derived Cancer Stem Cell LinesAna Teresa Serra0Margarida Serra1Ana Carina Silva2Tamara Brckalo3Anita Seshire4Catarina Brito5Michael Wolf6Paula M. Alves7Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, PortugalInstituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, PortugalInstituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, PortugalMerck KGaA, Merck Serono, ImmunoOncology, Emerging Immunotherapies, Frankfurter Str. 250, 64293 Darmstadt, GermanyMerck KGaA, Merck Serono, ImmunoOncology, Emerging Immunotherapies, Frankfurter Str. 250, 64293 Darmstadt, GermanyInstituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, PortugalMerck KGaA, Merck Serono, ImmunoOncology, Emerging Immunotherapies, Frankfurter Str. 250, 64293 Darmstadt, GermanyInstituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, PortugalCancer stem cells (CSCs) have recently raised great interest as a promising biological system for designing effective cancer therapies. The scarcity of CSCs in vivo and the consequent low numbers obtained from biopsies represent a major hurdle to the development of such strategies. It is therefore necessary to design robust scalable methods to enable efficient expansion of bona fide CSCs in vitro. Here, we evaluated the applicability of computer-controlled bioreactors combined with 3D aggregate culture and microcarrier technology, widely used in stem cell bioprocessing, for the expansion and enrichment of CSCs isolated from different types of solid tumors—colorectal cancer (CRC) and non-small-cell lung cancer (NSCLC) from two patients. Results show that these culture strategies improved cell expansion and CSC enrichment. Both patient-derived CSC lines were able to grow on microcarriers, the best results being achieved for PPlus 102-L, Pro-F 102-L, Fact 102-L, and CGEN 102-L beads (5-fold and 40-fold increase in total cell concentration for CRC and NSCLC cells, respectively, in 6 days). As for 3D aggregate culture strategy, the cell proliferation profile was donor dependent. NSCLC cells were the only cells able to form aggregates and proliferate, and the flat-bottom bioreactor vessel equipped with a trapezoid-shaped paddle impeller was the most efficient configuration for cell growth (21-fold increase in cell concentration achieved in 8 days). Serum-free medium promotes CSC enrichment in both 3D aggregate and microcarrier cultures. The protocols developed herein for CSC expansion have the potential to be transferred to clinical and industrial settings, providing key insights to guide bioprocess design towards the production of enriched CSC cultures in higher quantity and improved quality.http://dx.doi.org/10.1155/2019/8347595 |
| spellingShingle | Ana Teresa Serra Margarida Serra Ana Carina Silva Tamara Brckalo Anita Seshire Catarina Brito Michael Wolf Paula M. Alves Scalable Culture Strategies for the Expansion of Patient-Derived Cancer Stem Cell Lines Stem Cells International |
| title | Scalable Culture Strategies for the Expansion of Patient-Derived Cancer Stem Cell Lines |
| title_full | Scalable Culture Strategies for the Expansion of Patient-Derived Cancer Stem Cell Lines |
| title_fullStr | Scalable Culture Strategies for the Expansion of Patient-Derived Cancer Stem Cell Lines |
| title_full_unstemmed | Scalable Culture Strategies for the Expansion of Patient-Derived Cancer Stem Cell Lines |
| title_short | Scalable Culture Strategies for the Expansion of Patient-Derived Cancer Stem Cell Lines |
| title_sort | scalable culture strategies for the expansion of patient derived cancer stem cell lines |
| url | http://dx.doi.org/10.1155/2019/8347595 |
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