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Microdroplet Systems for Gene Transfer: From Fundamentals to Future Perspectives
(MDPI AG, 2025-10-31)
Criollo, Mishell
;
Gina Layedra
;
Camilo Pérez Sosa
;
Gustavo Rosero
;
PEÑAHERRERA PAZMIÑO, ANA BELÉN
Microfluidics enables precise control of fluid movement within microchannels, facilitating the generation of microdroplets at high frequencies. This technology provides a unique platform for conducting biological and chemical experiments, enhancing throughput and sensitivity, particularly in single-cell analysis. The microdroplet environment enhances interactions between cells and gene delivery materials, resulting in greater contact area, higher reagent concentration, and improved diffusion for both eukaryotic and prokaryotic cells. This review discusses the advantages and limitations of transfection and transformation within microdroplet technologies, highlighting their potential to improve gene editing efficiency while addressing challenges related to delivery mechanisms and cellular uptake rates. The integration of microdroplet technology with advanced gene editing tools, such as CRISPR/Cas9, promises to streamline processes and improve outcomes in various applications, including therapeutic interventions, vaccine development, regenerative medicine, and personalized medicine. These advancements could lead to more precise targeting of genetic modifications, resulting in tailored therapies that better meet individual patient needs. Overall, the integration of gene delivery in microdroplets represents a significant leap in biotechnology, enhancing the efficacy of gene delivery systems and opening new avenues for research and development in precision medicine.
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Activation and Expansion of Human T-Cells Using Microfluidic Devices
(MDPI AG, 2025-04-25)
PEÑAHERRERA PAZMIÑO, ANA BELÉN
;
Gustavo Rosero
;
Dario Ruarte
;
Julia Pinter
;
Vizuete, Karla
Treatment of cancer patients with autologous T-cells expressing a chimeric antigen receptor (CAR) is one of the most promising therapeutic modalities for hematological malignancy treatment. For this treatment, primary T-cell expansion is needed. Microfluidic technologies can be used to better understand T-cell activation and proliferation. Microfluidics have had a meaningful impact in the way experimental biology and biomedical research are approached in general. Furthermore, microfluidic technology allows the generation of large amounts of data and enables the use of image processing for analysis. However, one of the major technical hurdles involved in growing suspension cells under microfluidic conditions is their immobilization, to avoid washing them out of the microfluidic chip during medium renewal. In this work, we use a multilevel microfluidic chip to successfully capture and immobilize suspension cells. Jurkat cells and T-cells are isolated through traps to microscopically track their development and proliferation after activation over a period of 8 days. The T-cell area of four independent microchannels was compared and there is no statistically significant difference between them (ANOVA p-value = 0.976). These multilevel microfluidic chips provide a new method of studying T-cell activation.
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In vitro gut-models to elucidate how human milk oligosaccharides shape the gut microbiota
(Elsevier BV, 2025-10)
PEÑAHERRERA PAZMIÑO, ANA BELÉN
;
Gustavo Rosero
;
Mishell Criollo
;
Julio Patricio Peñaherrera
This review explores the important role of human milk oligosaccharides (HMOs). It highlights remarkable properties of HMOs as they act as decoys for disease-causing microbes and prevent their adhesion to the gut epithelium enabling its expulsion from the intestine. The role of HMOs has been researched by conventional methodologies, such as cellular cultures and animal experimentation, yielding significant understanding; however, they are constrained in their ability to accurately emulate the intricacies of human pathology and evoke ethical concerns. Herein, in vitro gut-models (IVGM) applied in studies to elucidate how HMOs shape microbiota are presented. The majority of studies related to HMOs effect on gut microbiota are performed in bioreactors. This review presents a concise overview of this rapidly advancing technology, a cost analysis, an examination of contemporary applications and potential challenges associated with their implementation, and proposes recommendations for future trajectories over the forthcoming decade.

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