Cryogel-based Injectable 3D Microcarrier Co-culture for Support of Hematopoietic Progenitor Niches.

Details

Serval ID
serval:BIB_3CE5A09E5F62
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Cryogel-based Injectable 3D Microcarrier Co-culture for Support of Hematopoietic Progenitor Niches.
Journal
Current protocols
Author(s)
Tavakol D.N., Bonini F., Tratwal J., Genta M., Brefie-Guth J., Braschler T., Naveiras O.
ISSN
2691-1299 (Electronic)
ISSN-L
2691-1299
Publication state
Published
Issued date
11/2021
Peer-reviewed
Oui
Volume
1
Number
11
Pages
e275
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
Although hematopoietic stem cell (HSC) transplantation can restore functional hematopoiesis upon immune or chemotherapy-induced bone marrow failure, complications often arise during recovery, leading to up to 25% transplant-related mortality in treated patients. In hematopoietic homeostasis and regeneration, HSCs in the bone marrow give rise to the entirety of cellular blood components. One of the challenges in studying hematopoiesis is the ability to successfully mimic the relationship between the stroma and hematopoietic stem and progenitor cells (HSPCs). This study and the described protocols propose an advantageous method for culturing and assessing stromal hematopoietic support in three dimensions, representing a simplified in vitro model of the bone marrow niche that can be transplanted in vivo by injection. By co-culturing OP9 bone marrow-derived stromal cells (BMSCs) and cKit <sup>+</sup> Sca-1 <sup>+</sup> Lin <sup>-</sup> (KLS <sup>+</sup> ) HSPCs on collagen-coated carboxymethylcellulose scaffolds for 2 weeks in the absence of cytokines, we established a methodology for in vivo subcutaneous transplantation. With this model we were able to detect early signs of extramedullary hematopoiesis. This work can be useful for studying various stromal cell populations in co-culture, as well as simple transfer by injection of these scaffolds in vivo for heterotopic regeneration of the marrow microenvironment. © 2021 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Isolation of HSPCs from mice Basic Protocol 2: Co-seeding of HSPCs and BMSCs on collagen-coated CCMs Basic Protocol 3: Maintenance, real-time imaging, and analysis of co-seeded scaffolds Basic Protocol 4: End-point analysis of co-seeded scaffolds using flow cytometry and CFU assays Basic Protocol 5: Transplantation of scaffolds by subcutaneous injection Support Protocol: Preparation of custom scaffold drying device.
Keywords
Animals, Coculture Techniques, Cryogels, Hematopoiesis, Hematopoietic Stem Cells, Humans, Mesenchymal Stem Cells, Mice, biomaterial, bone marrow, cryogel, hematopoietic, injectable, scaffold, stem cell niche, tissue engineering
Pubmed
Open Access
Yes
Create date
03/12/2021 12:29
Last modification date
11/12/2021 7:38
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