Benefits of cryopreservation as long-term storage method of encapsulated cardiosphere-derived cells for cardiac therapy: A biomechanical analysis

Paz-Artigas, Laura; Ziani, Kaoutar; Alcaine, Clara; Báez-Díaz, Claudia; Blanco-Blázquez, Virginia; Pedraz, José Luís; Ochoa, Ignacio; Ciriza, Jesús

doi:10.1016/j.ijpharm.2021.121014

Cited by 5 publications

(3 citation statements)

References 58 publications

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“… 9 To study spheroid stiffness, we applied a constriction assay methodology ( Figure 6 A–C) previously developed in our group for the mechanical characterization of alginate-based microcapsules. 26 , 27 Either 20.000 or 40.000 cell seeding spheroids were studied by this system. However, although 20.000 cell seeding spheroids were enough to generate spheroids over 500 μm, they were not retained in the constriction system, not allowing the quantification of the stiffness by this methodology ( Figure S4 ).…”

Section: Resultsmentioning

confidence: 99%

Generation of Self-Induced Myocardial Ischemia in Large-Sized Cardiac Spheroids without Alteration of Environmental Conditions Recreates Fibrotic Remodeling and Tissue Stiffening Revealed by Constriction Assays

Paz-Artigas,

González-Lana,

Polo

et al. 2024

ACS Biomater. Sci. Eng.

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A combination of human-induced pluripotent stem cells (hiPSCs) and 3D microtissue culture techniques allows the generation of models that recapitulate the cardiac microenvironment for preclinical research of new treatments. In particular, spheroids represent the simplest approach to culture cells in 3D and generate gradients of cellular access to the media, mimicking the effects of an ischemic event. However, previous models required incubation under low oxygen conditions or deprived nutrient media to recreate ischemia. Here, we describe the generation of large spheroids (i.e., larger than 500 μm diameter) that self-induce an ischemic core. Spheroids were generated by coculture of cardiomyocytes derived from hiPSCs (hiPSC-CMs) and primary human cardiac fibroblast (hCF). In the proper medium, cells formed aggregates that generated an ischemic core 2 days after seeding. Spheroids also showed spontaneous cellular reorganization after 10 days, with hiPSC-CMs located at the center and surrounded by hCFs. This led to an increase in microtissue stiffness, characterized by the implementation of a constriction assay. All in all, these phenomena are hints of the fibrotic tissue remodeling secondary to a cardiac ischemic event, thus demonstrating the suitability of these spheroids for the modeling of human cardiac ischemia and its potential application for new treatments and drug research.

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Section: Resultsmentioning

confidence: 99%

Generation of Self-Induced Myocardial Ischemia in Large-Sized Cardiac Spheroids without Alteration of Environmental Conditions Recreates Fibrotic Remodeling and Tissue Stiffening Revealed by Constriction Assays

Paz-Artigas,

González-Lana,

Polo

et al. 2024

ACS Biomater. Sci. Eng.

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“…Therefore, the microencapsulation of cells has broad application prospects. Regarding cryopreservation, β cells can be microencapsulated for cryopreservation and delivered to the body; cardiosphere-derived cells (CDCs) are microencapsulated and frozen to treat heart disease; and encapsulating and cryopreserving small amounts of sperm can be used for assisted reproduction technology . Not limited to cryopreservation, microencapsulation can also be used to deliver cardiomyocytes to treat cardiovascular disease or deliver cells and proteins to cure bone defects. ,, In addition, microencapsulation is only one form of cell encapsulation, and fiber or film encapsulation can also be achieved.…”

Section: Discussionmentioning

confidence: 99%

Hydrogel Microencapsulation Enhances Cryopreservation of Red Blood Cells with Trehalose

Yao

Shen

Chen

et al. 2022

ACS Biomater. Sci. Eng.

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Cryopreservation of red blood cells (RBCs) plays a vital role in preserving rare blood and serologic testing, which is essential for clinical transfusion medicine. The main difficulties of the current cryopreservation technique are the high glycerol concentration and the tedious deglycerolization procedure after thawing. In this study, we explored a microencapsulation method for cryopreservation. RBC− hydrogel microcapsules with a diameter of approximately 2.184 ± 0.061 mm were generated by an electrostatic spraying device. Then, 0.7 M trehalose was used as a cryoprotective agent (CPA), and microcapsules were adhered to a stainless steel grid for liquid nitrogen freezing. The results show that compared with the RBCs frozen by cryovials, the recovery of RBCs after microencapsulation is significantly improved, up to a maximum of more than 85%. Additionally, the washing process can be completed using only 0.9% NaCl. After washing, the RBCs maintained their morphology and adenosine 5′-triphosphate (ATP) levels and met clinical transfusion standards. The microencapsulation method provides a promising, referenceable, and more practical strategy for future clinical transfusion medicine.

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“…Furthermore, these methods cannot further increase the hematocrit of cryopreserved RBCs, otherwise the recovery will drop drastically [ 29 , 42 , 44 ]. Recently, the use of alginate hydrogels to encapsulate various cells for cryopreservation has attracted wide attention [ 45 – 52 ], for the reason that alginate hydrogel can facilitate the vitrification of water encapsulated in small hydrogel microcapsules at low concentrations of CPAs, and suppress the formation of fatal ice crystals in the warming process [ 53 , 54 ]. Our previous study has proved that hydrogel encapsulation could enhance the RBC cryopreservation with low concentrations of glycerol, although the recovery was not high (< 80%) [ 55 ].…”

Section: Introductionmentioning

confidence: 99%

Core–Shell Microfiber Encapsulation Enables Glycerol-Free Cryopreservation of RBCs with High Hematocrit

Qin,

Chen,

Shen

et al. 2023

Nano-Micro Lett.

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Cryopreservation of red blood cells (RBCs) provides great potential benefits for providing transfusion timely in emergencies. High concentrations of glycerol (20% or 40%) are used for RBC cryopreservation in current clinical practice, which results in cytotoxicity and osmotic injuries that must be carefully controlled. However, existing studies on the low-glycerol cryopreservation of RBCs still suffer from the bottleneck of low hematocrit levels, which require relatively large storage space and an extra concentration process before transfusion, making it inconvenient (time-consuming, and also may cause injury and sample lose) for clinical applications. To this end, we develop a novel method for the glycerol-free cryopreservation of human RBCs with a high final hematocrit by using trehalose as the sole cryoprotectant to dehydrate RBCs and using core–shell alginate hydrogel microfibers to enhance heat transfer during cryopreservation. Different from previous studies, we achieve the cryopreservation of human RBCs at high hematocrit (> 40%) with high recovery (up to 95%). Additionally, the washed RBCs post-cryopreserved are proved to maintain their morphology, mechanics, and functional properties. This may provide a nontoxic, high-efficiency, and glycerol-free approach for RBC cryopreservation, along with potential clinical transfusion benefits.

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Benefits of cryopreservation as long-term storage method of encapsulated cardiosphere-derived cells for cardiac therapy: A biomechanical analysis

Cited by 5 publications

References 58 publications

Generation of Self-Induced Myocardial Ischemia in Large-Sized Cardiac Spheroids without Alteration of Environmental Conditions Recreates Fibrotic Remodeling and Tissue Stiffening Revealed by Constriction Assays

Generation of Self-Induced Myocardial Ischemia in Large-Sized Cardiac Spheroids without Alteration of Environmental Conditions Recreates Fibrotic Remodeling and Tissue Stiffening Revealed by Constriction Assays

Hydrogel Microencapsulation Enhances Cryopreservation of Red Blood Cells with Trehalose

Core–Shell Microfiber Encapsulation Enables Glycerol-Free Cryopreservation of RBCs with High Hematocrit

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