A Microwell Device for the Efficient Generation of Arrays of Microtissues and Humanized Bone Marrow Micro-Ossicles
(1) Background: There are no high-throughput microtissue platforms for generating bone marrow micro-ossicles. Herein, we describe a method for the assembly of arrays of microtissues from bone marrow stromal cells (BMSC) in vitro and their maturation into bone marrow micro-ossicles in vivo. (2) Metho...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-06-01
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| Series: | Organoids |
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| Online Access: | https://www.mdpi.com/2674-1172/2/2/8 |
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| author | Kathryn Futrega Md. Shafiullah Shajib Pamela G. Robey Michael R. Doran |
| author_facet | Kathryn Futrega Md. Shafiullah Shajib Pamela G. Robey Michael R. Doran |
| author_sort | Kathryn Futrega |
| collection | DOAJ |
| description | (1) Background: There are no high-throughput microtissue platforms for generating bone marrow micro-ossicles. Herein, we describe a method for the assembly of arrays of microtissues from bone marrow stromal cells (BMSC) in vitro and their maturation into bone marrow micro-ossicles in vivo. (2) Methods: Discs with arrays of 50 microwells were used to assemble microtissues from 3 × 10<sup>5</sup> BMSCs each on a nylon mesh carrier. Microtissues were cultured in chondrogenic induction medium followed by hypertrophic medium in an attempt to drive endochondral ossification, and then they were implanted in NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice, where they were remodeled into bone marrow micro-ossicles. Mice were transplanted with 10<sup>5</sup> human umbilical cord blood CD34<sup>+</sup> cells. (3) Results: Micro-ossicles contained more human CD45<sup>+</sup> cells, but fewer human CD34<sup>+</sup> progenitor cells than mouse marrow. Human hematopoietic progenitor cells cycle rapidly at non-physiological rates in mouse marrow, and reduced CD34<sup>+</sup> cell content in micro-ossicles is consistent with the notion that the humanized niche better controls progenitor cell cycling. (4) Conclusions: Assembling microtissues in microwells, linked by a nylon membrane carrier, provides an elegant method to manufacture and handle arrays of microtissues with bone organ-like properties. More generally, this approach and platform could aid bridging the gap between in vitro microtissue manipulation and in vivo microtissue implantation. |
| first_indexed | 2024-03-11T02:03:58Z |
| format | Article |
| id | doaj.art-286154987fcb43ea9bce252b342a2b2b |
| institution | Directory Open Access Journal |
| issn | 2674-1172 |
| language | English |
| last_indexed | 2024-03-11T02:03:58Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Organoids |
| spelling | doaj.art-286154987fcb43ea9bce252b342a2b2b2023-11-18T11:58:52ZengMDPI AGOrganoids2674-11722023-06-012210211910.3390/organoids2020008A Microwell Device for the Efficient Generation of Arrays of Microtissues and Humanized Bone Marrow Micro-OssiclesKathryn Futrega0Md. Shafiullah Shajib1Pamela G. Robey2Michael R. Doran3National Institute of Dental and Craniofacial Research (NIDCR), National Institutes of Health (NIH), Department of Health and Human Services, Bethesda, MD 20892, USASchool of Biomedical Sciences, Queensland University of Technology, Brisbane 4000, AustraliaNational Institute of Dental and Craniofacial Research (NIDCR), National Institutes of Health (NIH), Department of Health and Human Services, Bethesda, MD 20892, USANational Institute of Dental and Craniofacial Research (NIDCR), National Institutes of Health (NIH), Department of Health and Human Services, Bethesda, MD 20892, USA(1) Background: There are no high-throughput microtissue platforms for generating bone marrow micro-ossicles. Herein, we describe a method for the assembly of arrays of microtissues from bone marrow stromal cells (BMSC) in vitro and their maturation into bone marrow micro-ossicles in vivo. (2) Methods: Discs with arrays of 50 microwells were used to assemble microtissues from 3 × 10<sup>5</sup> BMSCs each on a nylon mesh carrier. Microtissues were cultured in chondrogenic induction medium followed by hypertrophic medium in an attempt to drive endochondral ossification, and then they were implanted in NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice, where they were remodeled into bone marrow micro-ossicles. Mice were transplanted with 10<sup>5</sup> human umbilical cord blood CD34<sup>+</sup> cells. (3) Results: Micro-ossicles contained more human CD45<sup>+</sup> cells, but fewer human CD34<sup>+</sup> progenitor cells than mouse marrow. Human hematopoietic progenitor cells cycle rapidly at non-physiological rates in mouse marrow, and reduced CD34<sup>+</sup> cell content in micro-ossicles is consistent with the notion that the humanized niche better controls progenitor cell cycling. (4) Conclusions: Assembling microtissues in microwells, linked by a nylon membrane carrier, provides an elegant method to manufacture and handle arrays of microtissues with bone organ-like properties. More generally, this approach and platform could aid bridging the gap between in vitro microtissue manipulation and in vivo microtissue implantation.https://www.mdpi.com/2674-1172/2/2/8microwellmicrotissuemicro-ossiclebone marrowmesenchymal stem cellbone marrow stromal cell |
| spellingShingle | Kathryn Futrega Md. Shafiullah Shajib Pamela G. Robey Michael R. Doran A Microwell Device for the Efficient Generation of Arrays of Microtissues and Humanized Bone Marrow Micro-Ossicles Organoids microwell microtissue micro-ossicle bone marrow mesenchymal stem cell bone marrow stromal cell |
| title | A Microwell Device for the Efficient Generation of Arrays of Microtissues and Humanized Bone Marrow Micro-Ossicles |
| title_full | A Microwell Device for the Efficient Generation of Arrays of Microtissues and Humanized Bone Marrow Micro-Ossicles |
| title_fullStr | A Microwell Device for the Efficient Generation of Arrays of Microtissues and Humanized Bone Marrow Micro-Ossicles |
| title_full_unstemmed | A Microwell Device for the Efficient Generation of Arrays of Microtissues and Humanized Bone Marrow Micro-Ossicles |
| title_short | A Microwell Device for the Efficient Generation of Arrays of Microtissues and Humanized Bone Marrow Micro-Ossicles |
| title_sort | microwell device for the efficient generation of arrays of microtissues and humanized bone marrow micro ossicles |
| topic | microwell microtissue micro-ossicle bone marrow mesenchymal stem cell bone marrow stromal cell |
| url | https://www.mdpi.com/2674-1172/2/2/8 |
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