Long-Term Human Hematopoietic Stem Cell Culture in Microdroplets

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Carreras, Pilar and González, Itziar and Gallardo, Miguel and Ortiz-Ruiz, Alejandra and Morales, Maria Luz and Encinas, Jessica and Martínez López, Joaquín (2021) Long-Term Human Hematopoietic Stem Cell Culture in Microdroplets. Micromachines, 12 (1). p. 90. ISSN 2072-666X

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Official URL: https://doi.org/10.3390/mi12010090




Abstract

We previously reported a new approach for micromanipulation and encapsulation of human stem cells using a droplet-based microfluidic device. This approach demonstrated the possibility of encapsulating and culturing difficult-to-preserve primary human hematopoietic stem cells using an engineered double-layered bead composed by an inner layer of alginate and an outer layer of Puramatrix. We also demonstrated the maintenance and expansion of Multiple Myeloma cells in this construction. Here, the presented microfluidic technique is applied to construct a 3D biomimetic model to recapitulate the human hematopoietic stem cell niche using double-layered hydrogel beads cultured in 10% FBS culture medium. In this model, the long-term maintenance of the number of cells and expansion of hHSCS encapsulated in the proposed structures was observed. Additionally, a phenotypic characterization of the human hematopoietic stem cells generated in the presented biomimetic model was performed in order to assess their long-term stemness maintenance. Results indicate that the ex vivo cultured human CD34+ cells from bone marrow were viable, maintained, and expanded over a time span of eight weeks. This novel long-term stem cell culture methodology could represent a novel breakthrough to improve Hematopoietic Progenitor cell Transplant (HPT) as well as a novel tool for further study of the biochemical and biophysical factors influencing stem cell behavior. This technology opens a myriad of new applications as a universal stem cell niche model potentially able to expand other types of cells.


Item Type:Article
Uncontrolled Keywords:Microdroplets; Microfluidics; Stem cell culture; Hematopoietic stem cell
Subjects:Medical sciences > Medicine > Hematology
ID Code:67558
Deposited On:30 Aug 2021 08:19
Last Modified:06 Sep 2021 09:15

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