title: Sustained Administration of β-cell Mitogens to Intact Mouse Islets Ex Vivo Using Biodegradable Poly(lactic-co-glycolic acid) Microspheres.
abstract: The development of biomaterials has significantly increased the potential for targeted drug delivery to a variety of cell and tissue types, including the pancreatic β-cells. In addition, biomaterial particles, hydrogels, and scaffolds also provide a unique opportunity to administer sustained, controllable drug delivery to β-cells in culture and in transplanted tissue models. These technologies allow the study of candidate β-cell proliferation factors using intact islets and a translationally relevant system. Moreover, determining the effectiveness and feasibility of candidate factors for stimulating β-cell proliferation in a culture system is critical before moving forward to in vivo models. Herein, we describe a method to co-culture intact mouse islets with biodegradable compound of interest (COI)-loaded poly(lactic-co-glycolic acid) (PLGA) microspheres for the purpose of assessing the effects of sustained in situ release of mitogenic factors on β-cell proliferation. This technique describes in detail how to generate PLGA microspheres containing a desired cargo using commercially available reagents. While the described technique uses recombinant human Connective tissue growth factor (rhCTGF) as an example, a wide variety of COI could readily be used. Additionally, this method utilizes 96-well plates to minimize the amount of reagents necessary to assess β-cell proliferation. This protocol can be readily adapted to use alternative biomaterials and other endocrine cell characteristics such as cell survival and differentiation status.
authors.affiliation: Department of Medicine, Vanderbilt University Medical Center.
authors.forename: Raymond C
authors.initials: RC
authors.lastname: Pasek
authors.affiliation: School of Engineering, Vanderbilt University.
authors.forename: Taylor E
authors.initials: TE
authors.lastname: Kavanaugh
authors.affiliation: School of Engineering, Vanderbilt University.
authors.forename: Craig L
authors.initials: CL
authors.lastname: Duvall
authors.affiliation: Department of Medicine, Vanderbilt University Medical Center; Department of Veterans Affairs, Tennessee Valley Health Authority; maureen.gannon@vanderbilt.edu.
authors.forename: Maureen A
authors.initials: MA
authors.lastname: Gannon
mesh_terms: D000818:Animals; D001672:Biocompatible Materials; D018920:Coculture Techniques; D016503:Drug Delivery Systems; D006018:Glycols; D006801:Humans; D050417:Insulin-Secreting Cells; D007515:Islets of Langerhans; D019344:Lactic Acid; D051379:Mice; D008863:Microspheres; D008934:Mitogens; D011100:Polyglycolic Acid
chemical_list: D001672:Biocompatible Materials; D006018:Glycols; D008934:Mitogens; D011100:Polyglycolic Acid; D019344:Lactic Acid