Cell Encapsulation Research Articles

Postmodification with Polycations Enhances Key Properties of Alginate-Based Multicomponent Microcapsules.

Postmodification of alginate-based microspheres with polyelectrolytes (PEs) is commonly used in the cell encapsulation field to control microsphere stability and permeability. However, little is known about how different applied PEs shape the microsphere morphology and properties, particularly in vivo. Here, we addressed this question using model multicomponent alginate-based microcapsules postmodified with PEs of different charge and structure. We found that the postmodification can enhance or impair the mechanical resistance and biocompatibility of microcapsules implanted into a mouse model, with polycations surprisingly providing the best results. Confocal Raman microscopy and confocal laser scanning microscopy (CLSM) analyses revealed stable interpolyelectrolyte complex layers within the parent microcapsule, hindering the access of higher molar weight PEs into the microcapsule core. All microcapsules showed negative surface zeta potential, indicating that the postmodification PEs get hidden within the microcapsule membrane, which agrees with CLSM data. Human whole blood assay revealed complex behavior of microcapsules regarding their inflammatory and coagulation potential. Importantly, most of the postmodification PEs, including polycations, were found to be benign toward the encapsulated model cells.

Rapidly damping hydrogels engineered through molecular friction

Hydrogels capable of swift mechanical energy dissipation hold promise for a range of applications including impact protection, shock absorption, and enhanced damage resistance. Traditional energy absorption in such materials typically relies on viscoelastic mechanisms, involving sacrificial bond breakage, yet often suffers from prolonged recovery times. Here, we introduce a hydrogel designed for friction-based damping. This hydrogel features an internal structure that facilitates the motion of a chain walker within its network, effectively dissipating mechanical stress. The hydrogel network architecture allows for rapid restoration of its damping capacity, often within seconds, ensuring swift material recovery post-deformation. We further demonstrate that this hydrogel can significantly shield encapsulated cells from mechanical trauma under repetitive compression, owing to its proficient energy damping and rapid rebound characteristics. Therefore, this hydrogel has potential for dynamic load applications like artificial muscles and synthetic cartilage, expanding the use of hydrogel dampers in biomechanics and related areas.

Mixed κ/ι-carrageenan - LM pectin gels: Relating the rheological and mechanical properties with the capacity for probiotic encapsulation

The rheological and mechanical properties of mixed κ/ι-carrageenan - LM pectin gels were determined, and the potential of these gels for the formation of beads using the extrusion method and for the encapsulation of Lacticaseibacillus rhamnosus ATCC 53103 (LGG) was evaluated. Self-standing gels were obtained with all formulations evaluated. Carrageenan-rich gels, with carrageenan fraction (XC) ≥ 0.75, exhibited the highest storage modulus, but they were also brittle, while pectin-rich gels (XC ≤ 0.25) presented the highest hardness and cohesiveness. Pectin-rich formulations formed beads with the smallest initial diameter (2.40–2.45 mm), and the addition of carrageenan produced significantly more spherical beads compared to pure-pectin ones. As pectin-rich beads were the formulations that resisted simulated gastrointestinal conditions, these were selected for the encapsulation of LGG. These beads showed high encapsulation yields (87–96 %), and the percentage reduction of CFU/g during storage and simulated gastrointestinal conditions was not significantly different among formulations, the latter being significantly lower for encapsulated cells (8.64–15.03 %) compared to free cells (71.20 %). These results indicate that carrageenan-pectin gel beads with XC ≤ 0.25 were successful in encapsulating probiotic bacteria, and this capacity was related to the rheological and mechanical properties of the gels.

Bio-Inspired Screwed Conduits from the Microfluidic Rope-Coiling Effect for Microvessels and Bronchioles

Tubular microfibers have recently attracted extensive interest for applications in tissue engineering. However, the fabrication of tubular fibers with intricate hierarchical structures remains a major challenge. Here, we present a novel one-step microfluidic spinning method to generate bio-inspired screwed conduits (BSCs). Based on the microfluidic rope-coiling effect, a viscous hydrogel precursor is first curved into a helix stream in the channel, and then consecutively packed as a hollow structured stream and gelated into a screwed conduit (SC) via ionic and covalent crosslinking. By taking advantage of the excellent fluid-controlling ability of microfluidics, various tubes with diverse structures are fabricated via simple control over fluid velocities and multiple microfluidic device designs. The perfusability and permeability results, as well as the encapsulation and culture of human umbilical vein endothelial cells (HUVECs), human pulmonary alveolar epithelial cells (HPAs), and myogenic cells (C2C12), demonstrate that these SCs have good perfusability and permeability and the ability to induce the formation of functional biostructures. These features support the uniqueness and potential applications of these BSCs as biomimetic blood vessels and bronchiole tissues in combination with tissue microstructures, with likely application possibilities in biomedical engineering.

Advancing diagnostics and disease modeling: current concepts in biofabrication of soft microfluidic systems

Soft microfluidic systems play a pivotal role in personalized medicine, particularly in in vitro diagnostics tools and disease modeling. These systems offer unprecedented precision and versatility, enabling the creation of intricate three-dimensional (3D) tissue models that can closely emulate both physiological and pathophysiological conditions. By leveraging innovative biomaterials and bioinks, soft microfluidic systems can circumvent the current limitations involving the use of polydimethylsiloxane (PDMS), thus facilitating the development of customizable systems capable of sustaining the functions of encapsulated cells and mimicking complex biological microenvironments. The integration of lab-on-a-chip technologies with soft nanodevices further enhances disease models, paving the way for tailored therapeutic strategies. The current research concepts underscore the transformative potential of soft microfluidic systems, exemplified by recent breakthroughs in soft lithography and 3D (bio)printing. Novel applications, such as multi-layered tissues-on-chips and skin-on-a-chip devices, demonstrate significant advancements in disease modeling and personalized medicine. However, further exploration is warranted to address challenges in replicating intricate tissue structures while ensuring scalability and reproducibility. This exploration promises to drive innovation in biomedical research and healthcare, thus offering new insights and solutions to complex medical challenges and unmet needs.

Granular Biphasic Colloidal Hydrogels for 3D Bioprinting.

Granular hydrogels composed of hydrogel microparticles are promising candidates for 3D bioprinting due to their ability to protect encapsulated cells. However, to achieve high print fidelity, hydrogel microparticles need to jam to exhibit shear-thinning characteristics, which is crucial for 3D printing. Unfortunately, this overpacking can significantly impact cell viability, thereby negating the primary advantage of using hydrogel microparticles to shield cells from shear forces. To overcome this challenge, a novel solution: a biphasic, granular colloidal bioink designed to optimize cell viability and printing fidelity is introduced. The biphasic ink consists of cell-laden polyethylene glycol (PEG) hydrogel microparticles embedded in a continuous gelatin methacryloyl (GelMA)-nanosilicate colloidal network. Here, it is demonstrated that this biphasic bioink offers outstanding rheological properties, print fidelity, and structural stability. Furthermore, its utility for engineering complex tissues with multiple cell types and heterogeneous microenvironments is demonstrated, by incorporating β-islet cells into the PEG microparticles and endothelial cells in the GelMA-nanosilicate colloidal network. Using this approach, it is possible to induce cell patterning, enhance vascularization, and direct cellular function. The proposed biphasic bioink holds significant potential for numerous emerging biomedical applications, including tissue engineering and disease modeling.

Influence of Kluyveromyces lactis and Enterococcus faecalis on Obtaining Lactic Acid by Cheese Whey Fermentation

Cheese whey is a byproduct of the cheese industry that causes high levels of pollution in the environment, but its high lactose content means that it can be used as a source to obtain lactic acid. In this study, two strains, one belonging to a yeast and the other one to a bacteria (Kluyveromyces lactis and Enterococcus faecalis), were isolated from cheese whey and molecularly characterized, and the optimal growth conditions were determined. Then, using proteinized and deproteinized cheese whey, batch fermentation was carried out with the strains arranged in suspension and immobilized. The consumption of lactose and the production of lactic acid were measured through Brix degrees and acidity analysis. Afterwards, the lactic acid was purified, and its yield and physical and chemical characteristics were determined. It was proven that there were differences between each of the strains; arranged in free or encapsulated cells, the proteinized and deproteinized cheese wheys, under the same purification conditions, achieved different yields, colors, and densities of lactic acid. Immobilized Enterococcus faecalis had the highest yield (50.61 ± 34.94 g/L) using the deproteinized cheese whey compared to the immobilized Kluyveromyces lactis (35.70 ± 0.15 g/L) using the proteinized cheese whey.

Providing biomimetic microenvironment for pulp regeneration via hydrogel-mediated sustained delivery of tissue-specific developmental signals

Regenerative endodontic therapy is a promising approach to restore the vitality of necrotic teeth, however, pulp regeneration in mature permanent teeth remains a substantial challenge due to insufficient developmental signals. The dentin is embryologically and histologically similar to the pulp, which contains a cocktail of pulp-specific structural proteins and growth factors, thus we proposed an optimizing strategy to obtain dentin matrix extracted proteins (DMEP) and engineered a DMEP functionalized double network hydrogel, whose physicochemical property was tunable by adjusting polymer concentrations to synchronize with regenerated tissues. In vitro models showed that the biomimetic hydrogel with sustained release of DMEP provided a beneficial microenvironment for the encapsulation, propagation and migration of human dental pulp stem cells (hDPSCs). The odontogenic and angiogenic differentiation of hDPSCs were enhanced as well. To elicit the mechanism hidden in the microenvironment to guide cell fate, RNA sequencing was performed and 109 differential expression of genes were identified, the majority of which enriched in cell metabolism, cell differentiation and intercellular communications. The involvement of ERK, p38 and JNK MAPK signaling pathways in the process was confirmed. Of note, in vivo models showed that the injectable and in situ photo-crosslinkable hydrogel was user-friendly for root canal systems and was capable of inducing the regeneration of highly organized and vascularized pulp-like tissues in root segments that subcutaneously implanted into nude mice. Taken together, this study reported a facile and efficient way to fabricate a cell delivery hydrogel with pulp-specific developmental cues, which exhibited promising application and translation potential in future regenerative endodontic fields.

Spatial atomic layer deposition of nitrogen-doped alumina thin films for high-performance perovskite solar cell encapsulation

An atmospheric-pressure spatial atomic layer deposition (AP-SALD) system is used to deposit nitrogen-doped alumina (N-AlOx) thin-film-encapsulation layers. The rapid nature of the AP-SALD process facilitates deposition of 60-nm layers directly on perovskite solar cells at 130 °C with no damage to the temperature-sensitive perovskite and organic materials. Varying the bubbling of a NH4OH precursor varied the nitrogen concentration from 0.08 to 0.68 at%. These small concentrations were found to have a significant impact on the structural properties of the films and their moisture barrier performance. The N-AlOx thin films had slightly higher growth rates than undoped AlOx, less unwanted hydroxyl and carbon content, and were smoother and more compact, which was attributed to a higher flux of reactive species from the volatile NH4OH. Optical calcium tests showed that the N-AlOx films had lower water-vapor-transmission rates (∼10-5 g/m2/day) than undoped AlOx films and the transmission was minimized for 0.28% nitrogen. The increased compactness of the N-AlOx films is expected to minimize nanoscale percolation pathways, whereas higher nitrogen-defect concentrations may facilitate water permeation through these pathways. The stability of n-i-p and p-i-n perovskite solar cells under standard ISOS-D-1 and ISOS-D-3 testing conditions was significantly enhanced by the encapsulation layers. An N-AlOx encapsulation layer with 0.28% nitrogen improved the T80 value of a p-i-n formamidinium methylammonium lead iodide solar cell from 144 hrs to 855 hrs (ISOS-D-1) and 52 hrs to 300 hrs (ISOS-D-3).

Six‐Month Outdoor Performance Study of Stable Perovskite Solar Cells Under Real Operating Conditions

Perovskite solar cells (PSCs) have sparked great excitement in the photovoltaics community due to their remarkable efficiency, flexibility, and ability to be synthesized at low cost. However, their instability poses a major roadblock to their widespread adoption. Recognizing this challenge, this work describes the performance of stable PSCs operating under real‐life conditions in the Paris area. Two state‐of‐the‐art 2D/3D (HOOC(CH2)4NH3)2PbI4/CH3NH3PbI3 encapsulated cells are considered—one with maximum power point tracking and hourly current–voltage curve measurements, and another connected to a fixed resistive load. Their performance is compared over six months, and a detailed analysis is conducted on the cells’ hysteresis and electrical response to varying weather conditions. Although the first cell starts to degrade after a few months of operation, the second one remains remarkably stable, highlighting critical issues with outdoor tracking. Given that the stability of this PSC architecture has already been established under controlled standard conditions for over one year, these exciting findings pave the way for the commercialization of perovskite‐based photovoltaic devices.

Optogenetic therapeutic strategies for diabetes mellitus.

Diabetes mellitus (DM) is a common chronic disease affecting humans globally. It is characterized by abnormally elevated blood glucose levels due to the failure of insulin production or reduction of insulin sensitivity and functionality. Insulin and glucagon-like peptide (GLP)-1 replenishment or improvement of insulin resistance are the two major strategies to treat diabetes. Recently, optogenetics that uses genetically encoded light-sensitive proteins to precisely control cell functions has been regarded as a novel therapeutic strategy for diabetes. Here, we summarize the latest development of optogenetics and its integration with synthetic biology approaches to produce light-responsive cells for insulin/GLP-1 production, amelioration of insulin resistance and neuromodulation of insulin secretion. In addition, we introduce the development of cell encapsulation and delivery methods and smart bioelectronic devices for the in vivo application of optogenetics-based cell therapy in diabetes. The remaining challenges for optogenetics-based cell therapy in the clinical translational study are also discussed.

Impact of Silk-Ionomer Encapsulation on Immune Cell Mechanical Properties and Viability.

Encapsulation of single cells is a powerful technique used in various fields, such as regenerative medicine, drug delivery, tissue regeneration, cell-based therapies, and biotechnology. It offers a method to protect cells by providing cytocompatible coatings to strengthen cells against mechanical and environmental perturbations. Silk fibroin, derived from the silkworm Bombyx mori, is a promising protein biomaterial for cell encapsulation due to the cytocompatibility and capacity to maintain cell functionality. Here, THP-1 cells, a human leukemia monocytic cell line, were encapsulated with chemically modified silk polyelectrolytes through electrostatic layer-by-layer deposition. The effectiveness of the silk nanocoating was assessed using scanning electron microscopy (SEM) and confocal microscopy and on cell viability and proliferation by Alamar Blue assay and live/dead staining. An analysis of the mechanical properties of the encapsulated cells was conducted using atomic force microscopy nanoindentation to measure elasticity maps and cellular stiffness. After the cells were encapsulated in silk, an increase in their stiffness was observed. Based on this observation, we developed a mechanical predictive model to estimate the variations in stiffness in relation to the thickness of the coating. By tuning the cellular assembly and biomechanics, these encapsulations promote systems that protect cells during biomaterial deposition or processing in general.

Mathematical Evaluation of Population Changes of Lactobacillus acidophilus and Bifidobacterium animalis ssp. lactis as Free and Encapsulated Cells in Butter

Growing butter markets, domestically and globally, provide opportunities for value-added variants of butter. Adding probiotics to butter could boosts its bioactivity; however, maintaining probiotic viability during storage is a major challenge. Mathematical analysis of probiotic population changes could help improve our understanding of how probiotics interact with butter and storage conditions. Two strains of probiotics in a 1:1 ratio as free cells or Whey Protein Hydrolysate–Maltodextrin (WPH-MD)-encapsulated cells, Lactobacillus acidophilus ATCC 4356 (LA5) and Bifidobacterium animalis ssp. lactis ATCC 27536 (BB12), were separately mixed into butter at 1% levels. Using analysis of covariance, a mathematical evaluation for probiotic population changes was performed by periodically determining viable counts, resulting in an adjusted R2 value of 0.98 and demonstrating a strong relationship between the dependent variable (log10 counts of probiotics) and independent variables (cell type, temperature of storage, and time of storage). After 21 days of storage, the number of free cells in butter dropped from 7.45 log10 CFU/g to 0.56 log10 CFU/g. On the other hand, it took 63 days for encapsulated cells to achieve 0.80 log10 CFU/g at the same temperature. The same trend persisted at −18 °C, indicating that the WPH-MD encapsulant had a protective effect.

Calcium Alginate Core–Shell Liquid Beads Encapsulated with Microalgae for Wastewater Treatment

Liquid beads are core–shell particles with a liquid core and a solid shell. Calcium alginate liquid beads have been emerging as a promising platform for cell encapsulation. These beads have demonstrated their capability of encapsulating and culturing a wide range of human cells for tissue engineering. However, a significant research gap remains in the application of alginate liquid beads for encapsulating photosynthetic microorganisms. Herein, fast‐growing microalgae strain Chlorella vulgaris is encapsulated into calcium alginate liquid beads to facilitate the removal of nutrients from wastewater, minimizing the risk of eutrophication. Liquid bead‐microalgae systems are prepared, using different calcium ion concentrations as crosslinking ions. It has been thoroughly characterized for their morphologies, cell growth patterns, nutrient removal capabilities, and overall stability throughout the wastewater treatment process, with upflow anaerobic sludge blanket effluent as the wastewater model. The results indicate that the liquid bead‐microalgae system with the highest calcium ion concentration (5%) performs more efficiently, exhibiting a well‐formed crosslinking structure, leading to rapid cell growth with the highest cell density and the most effective removal of nutrients. The findings from this study provide valuable insights for future optimization and upscaling efforts in wastewater treatment systems based on calcium alginate liquid beads.

Improved perovskite photostability via application of TiO2, ZnO and AZO thin films by pulsed laser deposition

The UV-induced degradation of perovskite solar cells has been a barrier to commercialization. Thin film encapsulation represents a promising solution for extending perovskite solar cell lifetimes. Three materials with suitable bandgaps for blocking UV light are identified: TiO2, ZnO and AZO. Herein, the optical properties of TiO2, ZnO and AZO thin films grown by pulsed laser deposition at room-temperature are optimized by varying the oxygen partial pressure between 2, 5 and 20 mTorr during deposition. UV–Vis spectroscopy reveals facile bandgap tuning via the PLD O2 pressure for maximized UV light absorption/reflection. In particular, the TiO2 bandgap is found to narrow with decreasing O2 partial pressure as a result of an increase in oxygen vacancies and Ti3+ interstitials. Conversely, the AZO bandgap widens with decreasing O2 partial pressure, as explained by the Burstein-Moss effect. Finally, ZnO films have stable optical properties at all investigated oxygen pressures. The analysis finds that ZnO, TiO2, and AZO coatings exhibit maximum percentages of absorbed/reflected UV-A light of 72.13 ± 3.05%, 52.30 ± 2.72%, and 52.06 ± 2.20% at 5, 2 and 20 mTorr O2, respectively. Accelerated UV-aging experiments find that the films which screen the most UV light are also most effective in delaying perovskite degradation. A comparison of TiO2, ZnO and AZO photoprotective coatings concludes that ZnO is most suitable for perovskite solar cell encapsulation since it provides a ∼ 50% reduction in UV-degradation without significantly compromising visible light transmittance.

Characterizing an injectable alginate hydrogel as a co‐encapsulating system for beta cells and curcumin in type 1 diabetes therapy

AbstractEncapsulating insulin‐secreting cells within biocompatible hydrogels is a tissue engineering approach for type 1 diabetes treatment. Macroencapsulation is preferred over other encapsulation methods due to its easy implantation and retrieval. However, this treatment has challenges, including the need for invasive surgery for macrocapsule implantation and host response, resulting in fibrotic overgrowth around the implant and subsequent failure of the graft. Herein, an injectable alginate hydrogel, fabricated by different concentrations of as a retarding agent, was applied to avoid surgery. According to characterization tests, hydrogel made by 0.3 M showed a higher swelling ratio and diffusion coefficient, appropriate for the diffusion of nutrients, oxygen, and insulin. It was also stiffer with a lower swelling rate, demonstrating more robust elastic and solid‐like behaviour, suitable for cell attachment. Its small pore size (133.04 ± 53.28 μm), additionally, was essential for inhibiting immunoglobulins penetration. Thus, this hydrogel was used for RIN‐5F cells encapsulation and demonstrated good biocompatibility (>88% viability), and maintained the insulin‐releasing function of cells. Afterward, we co‐encapsulated beta cells with curcumin as an anti‐inflammatory drug and investigated the effect of this agent on cell behaviour in vitro. It was indicated that curcumin did not adversely affect viability and insulin secretion.

Single-Cell Screening through Cell Encapsulation in Photopolymerized Gelatin Methacryloyl

This study evaluated the potential of gelatin methacryloyl (GelMA) for single-cell screening compared to polyethylene glycol diacrylate (PEGDA). GelMA photopolymerized at 1000–2000 mJ/cm2 produced consistent patterns and supported HeLa cell viability. GelMA (5%w/v) facilitated better cell collection within 2 days due to its shape retention. GelMA demonstrated biocompatibility with HeLa cells exhibiting exponential proliferation and biodegradation over 5 days. The average cell displacement over 2 days was 16 µm. Two targeted cell recovery strategies using trypsin were developed: one for adherent cells encapsulated at 800 mJ/cm2, and another for floating cells encapsulated at 800 mJ/cm2, enabling the selective removal of unwanted cells. These findings suggest GelMA as a promising biomaterial for single-cell screening applications, offering advantages over PEGDA in cell encapsulation and targeted recovery.

Fully synthetic, tunable poly(α-amino acids) as the base of bioinks curable by visible light

Bioinks play a crucial role in tissue engineering, influencing mechanical and chemical properties of the printed scaffold as well as the behavior of encapsulated cells. Recently, there has been a shift from animal origin materials to their synthetic alternatives. In this context, we present here bioinks based on fully synthetic and biodegradable poly(α,L-amino acids) (PolyAA) as an alternative to animal-based gelatin methacrylate (Gel-Ma) bioinks. Additionally, we first reported the possibility of the visible light photoinitiated incorporation of the bifunctional cell adhesive RGD peptide into the PolyAA hydrogel matrix. The obtained hydrogels are shown to be cytocompatible, and their mechanical properties closely resemble those of gelatin methacrylate-based scaffolds. Moreover, combining the unique properties of PolyAA-based bioinks, the photocrosslinking strategy, and the use of droplet-based printing allows the printing of constructs with high shape fidelity and structural integrity from low-viscosity bioinks without using any sacrificial components. Overall, presented PolyAA-based materials are a promising and versatile toolbox that extends the range of bioinks for droplet bioprinting.

Cell-adhesive double network self-healing hydrogel capable of cell and drug encapsulation: New platform to construct biomimetic environment with bottom-up approach

This study presents the development and characterization of a novel double-network self-healing hydrogel based on N-carboxyethyl chitosan (CEC) and oxidized dextran (OD) with the incorporation of crosslinked collagen (CEC-OD/COL-GP) to enhance its biological and physicochemical properties. The hydrogel formed via dynamic imine bond formation exhibited efficient self-healing within 30 min, and a compressive modulus recovery of 92 % within 2 h. In addition to its self-healing ability, CEC-OD/COL-GP possesses unique physicochemical characteristics including transparency, injectability, and adhesiveness to various substrates and tissues. Cell encapsulation studies confirmed the biocompatibility and suitability of the hydrogel as a cell-culture scaffold, with the presence of a collagen network that enhances cell adhesion, spreading, long-term cell viability, and proliferation. Leveraging their unique properties, we engineered assemblies of self-healing hydrogel modules for controlled spatiotemporal drug delivery and constructed co-culture models that simulate angiogenesis in tumor microenvironments. Overall, the CEC-OD/COL-GP hydrogel is a versatile and promising material for biomedical applications, offering a bottom-up approach for constructing complex structures with self-healing capabilities, controlled drug release, and support for diverse cell types in 3D environments. This hydrogel platform has considerable potential for advancements in tissue engineering and therapeutic interventions.

Evaluation of Alginate Hydrogel Microstrands for Stromal Cell Encapsulation and Maintenance.

Mesenchymal stromal cells (MSCs) have displayed potential in regenerating organ function due to their anti-fibrotic, anti-inflammatory, and regenerative properties. However, there is a need for delivery systems to enhance MSC retention while maintaining their anti-fibrotic characteristics. This study investigates the feasibility of using alginate hydrogel microstrands as a cell delivery vehicle to maintain MSC viability and phenotype. To accommodate cell implantation needs, we invented a Syringe-in-Syringe approach to reproducibly fabricate microstrands in small numbers with a diameter of around 200 µm and a porous structure, which would allow for transporting nutrients to cells by diffusion. Using murine NIH 3T3 fibroblasts and primary embryonic 16 (E16) salivary mesenchyme cells as primary stromal cell models, we assessed cell viability, growth, and expression of mesenchymal and fibrotic markers in microstrands. Cell viability remained higher than 90% for both cell types. To determine cell number within the microstrands prior to in vivo implantation, we have further optimized the alamarBlue assay to measure viable cell growth in microstrands. We have shown the effect of initial cell seeding density and culture period on cell viability and growth to accommodate future stromal cell delivery and implantation. Additionally, we confirmed homeostatic phenotype maintenance for E16 mesenchyme cells in microstrands.