A thermoreversible polymer mediates controlled release of glial cell line-derived neurotrophic factor to enhance kidney regeneration.
Previously, we reported that human mesenchymal stem cells (hMSCs) that were cultivated in growing embryos differentiated in an appropriate developmental milieu, thereby facilitating the development of a functional renal unit. However, this approach required transfection with an adenovirus that expressed glial cell line-derived neurotrophic factor (GDNF) to enhance the development of hMSC-derived renal tissue, and safety issues restrict the clinical use of such viral vectors. To circumvent this problem, we tested an artificial polymer as a means to diffuse GDNF. This GDNF-polymer, which exists in liquid form at 4 degrees C but becomes a hydrogel upon heating to 37 degrees C, was used as a thermoreversible switch, allowing the injection of hMSCs at low viscosity using a mouth pipette, with subsequent slow diffusion of GDNF as it solidified. The polymer, which was dissolved in a solution of GDNF at 4 degrees C and then maintained at 37 degrees C, acted as a diffuser of GDNF for more than 48 h. LacZ-transfected hMSCs and the GDNF-polymer (at 4 degrees C) were placed in the nephrogenic sites of growing rat embryos that were maintained at 37 degrees C. Forty-eight hours later, the resultant kidney anlagen were dissected out and allowed to continue developing for 6 days in vitro. Whole-organ X-Gal staining and fluorescence activated cell sorter analysis showed that the number of hMSC-derived cells was significantly increased in developed anlagen that have been generated from hMSCs plus GDNF-polymer compared with those from hMSCs plus GDNF-containing medium and was comparable to those from adenovirus-transfected hMSCs. These findings suggest that the GDNF-polymer can be used as a diffuser of GDNF for kidney organogenesis.
Gheisari
Yousof
Y
Project Laboratory for Kidney Regeneration, Institute of DNA Medicine, The Jikei University School of Medicine, Tokyo, Japan.
Yokoo
Takashi
T
Matsumoto
Kei
K
Fukui
Akira
A
Sugimoto
Naomi
N
Ohashi
Toya
T
Kawamura
Tetsuya
T
Hosoya
Tatsuo
T
Kobayashi
Eiji
E
D000818:Animals; D019556:COS Cells; D002454:Cell Differentiation; D002522:Chlorocebus aethiops; D005260:Female; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000008:administration & dosage*; D007267:Injections; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods*; D011092:Polyethylene Glycols / Q000627:therapeutic use*; D011109:Polymethacrylic Acids / Q000627:therapeutic use*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012038:Regeneration; D051437:Renal Insufficiency / Q000628:therapy*; D055471:Thermal Diffusion
C495039:GDNF protein, human; D051100:Glial Cell Line-Derived Neurotrophic Factor; D011109:Polymethacrylic Acids; C084450:poly(N-isopropylacrylamide-co-butyl methacrylate); D011092:Polyethylene Glycols