Metabolic switching, growth kinetics and cell yields in the scalable manufacture of stem cell-derived insulin-producing cells
Abstract Background Diabetes is a disease affecting over 500 million people globally due to insulin insufficiency or insensitivity. For individuals with type 1 diabetes, pancreatic islet transplantation can help regulate their blood glucose levels. However, the scarcity of cadaveric donor islets lim...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2024-01-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13287-023-03574-3 |
| _version_ | 1797363625368748032 |
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| author | Diepiriye G. Iworima Robert K. Baker Cara Ellis Chris Sherwood Lisa Zhan Alireza Rezania James M. Piret Timothy J. Kieffer |
| author_facet | Diepiriye G. Iworima Robert K. Baker Cara Ellis Chris Sherwood Lisa Zhan Alireza Rezania James M. Piret Timothy J. Kieffer |
| author_sort | Diepiriye G. Iworima |
| collection | DOAJ |
| description | Abstract Background Diabetes is a disease affecting over 500 million people globally due to insulin insufficiency or insensitivity. For individuals with type 1 diabetes, pancreatic islet transplantation can help regulate their blood glucose levels. However, the scarcity of cadaveric donor islets limits the number of people that could receive this therapy. To address this issue, human pluripotent stem cells offer a potentially unlimited source for generating insulin-producing cells through directed differentiation. Several protocols have been developed to make stem cell-derived insulin-producing cells. However, there is a lack of knowledge regarding the bioprocess parameters associated with these differentiation protocols and how they can be utilized to increase the cell yield. Methods We investigated various bioprocess parameters and quality target product profiles that may influence the differentiation pipeline using a seven-stage protocol in a scalable manner with CellSTACKs and vertical wheel bioreactors (PBS-Minis). Results Cells maintained > 80% viability through all stages of differentiation and appropriately expressed stage-specific markers. During the initial four stages leading up to the development of pancreatic progenitors, there was an increase in cell numbers. Following pancreatic progenitor stage, there was a gradual decrease in the percentage of proliferative cells, as determined by Ki67 positivity, and a significant loss of cells during the period of endocrine differentiation. By minimizing the occurrence of aggregate fusion, we were able to enhance cell yield during the later stages of differentiation. We suggest that glucose utilization and lactate production are cell quality attributes that should be considered during the characterization of insulin-producing cells derived from stem cells. Our findings also revealed a gradual metabolic shift from glycolysis, during the initial four stages of pancreatic progenitor formation, to oxidative phosphorylation later on during endocrine differentiation. Furthermore, the resulting insulin-producing cells exhibited a response to several secretagogues, including high glucose. Conclusion This study demonstrates process parameters such as glucose consumption and lactate production rates that may be used to facilitate the scalable manufacture of stem cell-derived insulin-producing cells. |
| first_indexed | 2024-03-08T16:23:51Z |
| format | Article |
| id | doaj.art-eef5378a3b1e42df99f87e9d84023768 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-03-08T16:23:51Z |
| publishDate | 2024-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-eef5378a3b1e42df99f87e9d840237682024-01-07T12:14:19ZengBMCStem Cell Research & Therapy1757-65122024-01-0115112710.1186/s13287-023-03574-3Metabolic switching, growth kinetics and cell yields in the scalable manufacture of stem cell-derived insulin-producing cellsDiepiriye G. Iworima0Robert K. Baker1Cara Ellis2Chris Sherwood3Lisa Zhan4Alireza Rezania5James M. Piret6Timothy J. Kieffer7Department of Cellular and Physiological Sciences, Life Sciences Institute, The University of British ColumbiaDepartment of Cellular and Physiological Sciences, Life Sciences Institute, The University of British ColumbiaDepartment of Cellular and Physiological Sciences, Life Sciences Institute, The University of British ColumbiaMichael Smith Laboratories, The University of British ColumbiaDepartment of Cellular and Physiological Sciences, Life Sciences Institute, The University of British ColumbiaCRISPR TherapeuticsSchool of Biomedical Engineering, The University of British ColumbiaDepartment of Cellular and Physiological Sciences, Life Sciences Institute, The University of British ColumbiaAbstract Background Diabetes is a disease affecting over 500 million people globally due to insulin insufficiency or insensitivity. For individuals with type 1 diabetes, pancreatic islet transplantation can help regulate their blood glucose levels. However, the scarcity of cadaveric donor islets limits the number of people that could receive this therapy. To address this issue, human pluripotent stem cells offer a potentially unlimited source for generating insulin-producing cells through directed differentiation. Several protocols have been developed to make stem cell-derived insulin-producing cells. However, there is a lack of knowledge regarding the bioprocess parameters associated with these differentiation protocols and how they can be utilized to increase the cell yield. Methods We investigated various bioprocess parameters and quality target product profiles that may influence the differentiation pipeline using a seven-stage protocol in a scalable manner with CellSTACKs and vertical wheel bioreactors (PBS-Minis). Results Cells maintained > 80% viability through all stages of differentiation and appropriately expressed stage-specific markers. During the initial four stages leading up to the development of pancreatic progenitors, there was an increase in cell numbers. Following pancreatic progenitor stage, there was a gradual decrease in the percentage of proliferative cells, as determined by Ki67 positivity, and a significant loss of cells during the period of endocrine differentiation. By minimizing the occurrence of aggregate fusion, we were able to enhance cell yield during the later stages of differentiation. We suggest that glucose utilization and lactate production are cell quality attributes that should be considered during the characterization of insulin-producing cells derived from stem cells. Our findings also revealed a gradual metabolic shift from glycolysis, during the initial four stages of pancreatic progenitor formation, to oxidative phosphorylation later on during endocrine differentiation. Furthermore, the resulting insulin-producing cells exhibited a response to several secretagogues, including high glucose. Conclusion This study demonstrates process parameters such as glucose consumption and lactate production rates that may be used to facilitate the scalable manufacture of stem cell-derived insulin-producing cells.https://doi.org/10.1186/s13287-023-03574-3Bioprocess developmentCell yieldPluripotent stem cellsDiabetesBeta cellsIslets |
| spellingShingle | Diepiriye G. Iworima Robert K. Baker Cara Ellis Chris Sherwood Lisa Zhan Alireza Rezania James M. Piret Timothy J. Kieffer Metabolic switching, growth kinetics and cell yields in the scalable manufacture of stem cell-derived insulin-producing cells Stem Cell Research & Therapy Bioprocess development Cell yield Pluripotent stem cells Diabetes Beta cells Islets |
| title | Metabolic switching, growth kinetics and cell yields in the scalable manufacture of stem cell-derived insulin-producing cells |
| title_full | Metabolic switching, growth kinetics and cell yields in the scalable manufacture of stem cell-derived insulin-producing cells |
| title_fullStr | Metabolic switching, growth kinetics and cell yields in the scalable manufacture of stem cell-derived insulin-producing cells |
| title_full_unstemmed | Metabolic switching, growth kinetics and cell yields in the scalable manufacture of stem cell-derived insulin-producing cells |
| title_short | Metabolic switching, growth kinetics and cell yields in the scalable manufacture of stem cell-derived insulin-producing cells |
| title_sort | metabolic switching growth kinetics and cell yields in the scalable manufacture of stem cell derived insulin producing cells |
| topic | Bioprocess development Cell yield Pluripotent stem cells Diabetes Beta cells Islets |
| url | https://doi.org/10.1186/s13287-023-03574-3 |
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