Umbilical cord mesenchymal stromal cells—from bench to bedside

Chetty, Shashank; Yarani, Reza; Swaminathan, Ganesh; Primavera, Rosita; Regmi, Shobha; Rai, Sravanthi; Zhong, Jim; Ganguly, Abantika; Thakor, Avnesh S.

doi:10.3389/fcell.2022.1006295

Cited by 15 publications

(7 citation statements)

References 145 publications

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“…Currently, WJ-MSCs have been extensively utilized in clinical trials [11,12,39], and to meet the standards for their clinical use, a GMP-compliant manufacturing process has been documented. Studies have outlined bioprocesses for WJ-MSC production, including the establishment of MCB, WCB, and DP [27], and comparisons with BM-MSCs for identity, safety, and function [28].…”

Section: Discussionmentioning

confidence: 99%

A GMP-compliant manufacturing method for Wharton’s jelly-derived mesenchymal stromal cells

Chu,

Zhang,

Zeng

et al. 2024

Stem Cell Res Ther

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Background Wharton’s jelly-derived mesenchymal stem cells (WJ-MSCs) hold great therapeutic potential in regenerative medicine. Therefore, it is crucial to establish a Good Manufacturing Practice (GMP)-compliant methodology for the isolation and culture of WJ-MSCs. Through comprehensive research, encompassing laboratory-scale experiments to pilot-scale studies, we aimed to develop standardized protocols ensuring the high yield and quality of WJ-MSCs manufacturing. Methods Firstly, optimization of parameters for the enzymatic digestion method used to isolate WJ-MSCs was conducted. These parameters included enzyme concentrations, digestion times, seeding densities, and culture media. Additionally, a comparative analysis between the explant method and the enzymatic digestion method was performed. Subsequently, the consecutive passaging of WJ-MSCs, specifically up to passage 9, was evaluated using the optimized method. Finally, manufacturing processes were developed and scaled up, starting from laboratory-scale flask-based production and progressing to pilot-scale cell factory-based production. Furthermore, a stability study was carried out to assess the storage and use of drug products (DPs). Results The optimal parameters for the enzymatic digestion method were a concentration of 0.4 PZ U/mL Collagenase NB6 and a digestion time of 3 h, resulting in a higher yield of P0 WJ-MSCs. In addition, a positive correlation between the weight of umbilical cord tissue and the quantities of P0 WJ-MSCs has been observed. Evaluation of different concentrations of human platelet lysate revealed that 2% and 5% concentrations resulted in similar levels of cell expansion. Comparative analysis revealed that the enzymatic digestion method exhibited faster outgrowth of WJ-MSCs compared to the explant method during the initial passage. Passages 2 to 5 exhibited higher viability and proliferation ability throughout consecutive passaging. Moreover, scalable manufacturing processes from the laboratory scale to the pilot scale were successfully developed, ensuring the production of high-quality WJ-MSCs. Multiple freeze-thaw cycles of the DPs led to reduced cell viability and viable cell concentration. Subsequent thawing and dilution of the DPs resulted in a significant decrease in both metrics, especially when stored at 20–27 °C. Conclusion This study offers valuable insights into optimizing the isolation and culture of WJ-MSCs. Our scalable manufacturing processes facilitate the large-scale production of high-quality WJ-MSCs. These findings contribute to the advancement of WJ-MSCs-based therapies in regenerative medicine.

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Section: Discussionmentioning

confidence: 99%

A GMP-compliant manufacturing method for Wharton’s jelly-derived mesenchymal stromal cells

Chu,

Zhang,

Zeng

et al. 2024

Stem Cell Res Ther

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“…Currently, UC-MSCs have been extensively utilized in clinical trials [11,12,34]. However, there has been limited focus on developing SOPs for GMP to ensure consistent quality of these cells.…”

Section: Discussionmentioning

confidence: 99%

Development of a GMP-Compliant Separation Method for Isolating Wharton's Jelly Derived Mesenchymal Stromal Cells from the Umbilical Cord

Chu,

Zhang,

Zeng

et al. 2023

Preprint

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Background Wharton's jelly derived mesenchymal stem cells (UC-MSCs) hold great therapeutic potential in regenerative medicine. However, GMP-compliant optimal methods for isolating UC-MSCs from UC tissue are still lacking. Additionally, there is a dearth of detailed research spanning from laboratory-scale to pilot-scale studies. Therefore, it is essential to establish standardized protocols that ensure the reproducibility and safety of UC-MSC manufacturing. Methods In this study, we aimed to explore and optimize parameters for the enzymatic digestion method used for isolating UC-MSCs. These parameters included enzyme concentrations, digestion times, seeding densities, and culture media. Additionally, we conducted a comparative analysis between the explant method and enzymatic digestion method. Subsequently, we evaluated the consecutive passaging stability of UC-MSCs, specifically up to passage 9, using the optimized enzymatic digestion method. Finally, we developed and scaled up manufacturing processes, starting from laboratory-scale flask-based production and progressing to pilot-scale cell factory-based production. Results The optimal parameters for the enzymatic digestion method were determined to be a concentration of 0.4 PZ U/mL Collagenase NB6 and a digestion time of 3 hours, resulting in a higher quantity of P0 UC-MSCs. Additionally, we observed a positive correlation between the initial cell seeding density and the number of P0 UC-MSCs. Evaluation of different concentrations of human platelet lysate (hPL) revealed that 2% and 5% concentrations resulted in similar levels of cell expansion, whereas a 10% concentration led to decreased cell expansion. Comparative analysis revealed that the enzymatic digestion method exhibited faster outgrowth of UC-MSCs compared to the explant method. However, after subsequent passages, there were no significant differences between the explant and enzymatic digestion methods in terms of cell proliferation, cell viability, and immunophenotype. Notably, consecutive passaging of UC-MSCs using the enzymatic digestion method demonstrated stability, with maintained cellular characteristics and functionality. Passages 2 to 5 exhibited higher viability and proliferation ability. Moreover, we successfully developed scalable manufacturing processes from the laboratory scale to the pilot scale, ensuring consistent production of high-quality UC-MSCs. Conclusion Our study provides valuable insights into the optimization of UC tissue processing protocols, the parameters for the enzymatic digestion method, and the comparative analysis of different isolation methods. We also demonstrated the stability of consecutive passaging using this method. Moreover, our scalable manufacturing processes enable large-scale production of high-quality UC-MSCs. These findings contribute to the advancement of UC-MSC-based therapies in regenerative medicine.

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“…MSCs can be harvested from various tissue types such as bone marrow, adipose tissue, umbilical cord, and blood [ 8 ]. UC-MSCs exhibit higher proliferative capacity and cell viability than do bone marrow-derived MSCs (BM-MSCs) and adipose tissue-derived MSCs (AD-MSCs) [ [9] , [10] , [11] , [12] , [13] , [14] ]. Furthermore, UC-MSCs exhibited higher levels of secreted therapeutic cytokines and a higher proliferative capacity than that of BM-MSCs and AD-MSCs [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

Harvesting methods of umbilical cord-derived mesenchymal stem cells from culture modulate cell properties and functions

Nakao,

Nagase

2024

Regenerative Therapy

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Umbilical cord mesenchymal stromal cells—from bench to bedside

Cited by 15 publications

References 145 publications

A GMP-compliant manufacturing method for Wharton’s jelly-derived mesenchymal stromal cells

A GMP-compliant manufacturing method for Wharton’s jelly-derived mesenchymal stromal cells

Development of a GMP-Compliant Separation Method for Isolating Wharton's Jelly Derived Mesenchymal Stromal Cells from the Umbilical Cord

Harvesting methods of umbilical cord-derived mesenchymal stem cells from culture modulate cell properties and functions

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