2020Biochemical Engineering JournalOpen access

Mesenchymal stromal cells maintain the major quality attributes when expanded in different bioreactor systems

Juliana de Sá da Silva, Patrícia Severino, T. I. Wodewotzky, Dimas Tadeu Covas, Kamilla Swiech, Luciana Cavalheiro Marti, Cláudio Alberto Torres Suazo

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Abstract

Mesenchymal Stromal Cells (MSCs) are being targeted for diverse cell-therapies applications. The MSC expansion in bioreactor is necessary to achieve the cell number required for clinical applications in accordance with cGMP guidelines. Since the culture conditions may influence the post-expansion MSC outcomes, in this work, human Umbilical Cord Matrix derived Mesenchymal Stromal Cells (hUCM-MSCs) were evaluated during and after expansion using CultiSpher-S® microcarrier with different bioreactor hydrodynamics: the single-use Wave-Induced Motion Bioreactor (WIMB) with a modified cellbag design proposed by our laboratory, and the conventional spinner flask. WIMB cultures showed longer cell culture times and lower cell yields than spinner flask culture, positioning the WIMB as a less suitable bioreactor than conventional stirred-based systems for high-yield MSC expansion bioprocess. Regarding cell biological functions, hUCM-MSCs harvested from the CultiSpher-S®/WIMB and the CultiSpher-S®/spinner kept determining major MSC molecular and biological characteristics according to immunophenotyping and multilineage differentiation assays. Moreover, biological processes were not regulated differently when hUCM-MSCs were cultivated in WIMB or spinner flask, indicating that post-expansion hUCM-MSCs characteristics are similar in both bioreactor systems.

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What this paper is about

Mesenchymal Stromal Cells (MSCs) are being targeted for diverse cell-therapies applications. The MSC expansion in bioreactor is necessary to achieve the cell number required for clinical applications in accordance with cGMP guidelines. Since the culture conditions may influence the post-expansion MSC outcomes, in this work, human Umbilical Cord Matrix derived Mesenchymal Stromal Cells (hUCM-MSCs) were evaluated during and after expansion using CultiSpher-S® microcarrier with different bioreactor hydrodynamics: the single-use Wave-Induced Motion Bioreactor (WIMB) with a modified cellbag design proposed by our laboratory, and the conventional spinner flask. WIMB cultures showed longer cell culture times and lower cell yields than spinner flask culture, positioning the WIMB as a less suitable bioreactor than conventional stirred-based systems for high-yield MSC expansion bioprocess. Regarding cell biological functions, hUCM-MSCs harvested from the CultiSpher-S®/WIMB and the CultiSpher-S®/spinner kept determining major MSC molecular and biological characteristics according to immunophenotyping and multilineage differentiation assays. Moreover, biological processes were not regulated differently when hUCM-MSCs were cultivated in WIMB or spinner flask, indicating that post-expansion hUCM-MSCs characteristics are similar in both bioreactor systems.

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Available abstract

Mesenchymal Stromal Cells (MSCs) are being targeted for diverse cell-therapies applications. The MSC expansion in bioreactor is necessary to achieve the cell number required for clinical applications in accordance with cGMP guidelines. Since the culture conditions may influence the post-expansion MSC outcomes, in this work, human Umbilical Cord Matrix derived Mesenchymal Stromal Cells (hUCM-MSCs) were evaluated during and after expansion using CultiSpher-S® microcarrier with different bioreactor hydrodynamics: the single-use Wave-Induced Motion Bioreactor (WIMB) with a modified cellbag design proposed by our laboratory, and the conventional spinner flask. WIMB cultures showed longer cell culture times and lower cell yields than spinner flask culture, positioning the WIMB as a less suitable bioreactor than conventional stirred-based systems for high-yield MSC expansion bioprocess. Regarding cell biological functions, hUCM-MSCs harvested from the CultiSpher-S®/WIMB and the CultiSpher-S®/spinner kept determining major MSC molecular and biological characteristics according to immunophenotyping and multilineage differentiation assays. Moreover, biological processes were not regulated differently when hUCM-MSCs were cultivated in WIMB or spinner flask, indicating that post-expansion hUCM-MSCs characteristics are similar in both bioreactor systems.

Key concepts: Microcarrier, Mesenchymal stem cell, Bioreactor, Bioprocess, Stromal cell, Cell biology, Cell, Biotechnology

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