Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Research advances in cryopreserved preparations of mesenchymal stem cells: technical innovations, application challenges, and quality control

doi: 10.3389/fbioe.2026.1717539

Figure Lengend Snippet: Workflow for creating an “off-the-shelf” MSCs therapy. MSCs are isolated from tissues like bone marrow or umbilical cord and expanded under GMP. A key step is cryopreservation, shown here with two advanced methods: (A) using novel DMSO-free CPAs like trehalose-based polymers, and (B) preserving MSCs in their native tissue niche. After thawing, cells must pass several critical quality control (QC) tests. These confirm the cells’ identity (e.g., via transcriptomic signature), ensure their safety (free of contaminants), and measure their potency for the specific therapy, using assays like MLR for immunomodulation or MPS for vasculogenesis.

Article Snippet: DMSO/Human Platelet Lysate (HPL)-Based , 5%–10% DMSO + Human Platelet Lysate (HPL) , • Xeno-free, higher clinical safety • HPL provides rich growth factors and adhesion proteins, aiding post-thaw recovery • A mainstream clinical-grade alternative to FBS. , • Batch-to-batch variability persists with HPL • DMSO toxicity remains a concern • Requires rigorous HPL donor screening and pathogen inactivation , • Develop chemically defined HPL mimetics or recombinant protein formulations to fundamentally address batch variability • Combine HPL with novel non-penetrating CPAs (e.g., trehalose derivatives) to aim for DMSO concentrations below 5%, reducing toxicity while maintaining high cell recovery , , .

Techniques: Isolation, Preserving, Control

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Research advances in cryopreserved preparations of mesenchymal stem cells: technical innovations, application challenges, and quality control

doi: 10.3389/fbioe.2026.1717539

Figure Lengend Snippet: The evolving paradigm of MSC cryopreservation: from traditional DMSO + FBS-based cryoprotectants to novel cryoprotective agents, native tissue niches, and vitrification cryopreservation, extending toward promising horizons such as automated manufacturing platform.

Article Snippet: DMSO/Human Platelet Lysate (HPL)-Based , 5%–10% DMSO + Human Platelet Lysate (HPL) , • Xeno-free, higher clinical safety • HPL provides rich growth factors and adhesion proteins, aiding post-thaw recovery • A mainstream clinical-grade alternative to FBS. , • Batch-to-batch variability persists with HPL • DMSO toxicity remains a concern • Requires rigorous HPL donor screening and pathogen inactivation , • Develop chemically defined HPL mimetics or recombinant protein formulations to fundamentally address batch variability • Combine HPL with novel non-penetrating CPAs (e.g., trehalose derivatives) to aim for DMSO concentrations below 5%, reducing toxicity while maintaining high cell recovery , , .

Techniques:

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Research advances in cryopreserved preparations of mesenchymal stem cells: technical innovations, application challenges, and quality control

doi: 10.3389/fbioe.2026.1717539

Figure Lengend Snippet: Mechanisms of cryoinjury in MSCs and a comparison of advanced protective strategies. This figure illustrates three key concepts in MSCs cryopreservation. On the left, the process of conventional cryopreservation is shown, where the use of agents like DMSO can still lead to significant cell damage from extracellular ice crystal formation (causing osmotic stress) and intracellular ice formation (causing mechanical rupture), in addition to direct cytotoxicity. In the center, the protective mechanism of novel, advanced CPAs like trehalose-based or zwitterionic polymers is depicted. These agents mitigate cryoinjury by inhibiting ice recrystallization (IRI), stabilizing the cell membrane, and balancing osmotic shifts without inherent toxicity. On the right, the concept of preserving MSCs within their native tissue niche is shown. The natural extracellular matrix (ECM) acts as a protective scaffold, physically buffering against stresses and maintaining the cellular microenvironment, which enhances post-thaw viability and recovery.

Article Snippet: DMSO/Human Platelet Lysate (HPL)-Based , 5%–10% DMSO + Human Platelet Lysate (HPL) , • Xeno-free, higher clinical safety • HPL provides rich growth factors and adhesion proteins, aiding post-thaw recovery • A mainstream clinical-grade alternative to FBS. , • Batch-to-batch variability persists with HPL • DMSO toxicity remains a concern • Requires rigorous HPL donor screening and pathogen inactivation , • Develop chemically defined HPL mimetics or recombinant protein formulations to fundamentally address batch variability • Combine HPL with novel non-penetrating CPAs (e.g., trehalose derivatives) to aim for DMSO concentrations below 5%, reducing toxicity while maintaining high cell recovery , , .

Techniques: Comparison, Recrystallization, Membrane, Preserving