Alginate microencapsulation technology for the percutaneous delivery of adipose-derived stem cells.
BACKGROUND
Autologous fat is the ideal soft-tissue filler; however, its widespread application is limited because of variable clinical results and poor survival. Engineered fillers have the potential to maximize survival. Alginate is a hydrogel copolymer that can be engineered into spheres of <200 μm, thus facilitating mass transfer, allowing for subcutaneous injection, and protecting cells from shearing forces.


METHODS
Alginate powder was dissolved in saline, and adipose-derived stem cells (ADSCs) were encapsulated (1 million cells/mL) in alginate using an electrostatic bead generator. To assess effects of injection on cell viability, microspheres containing ADSCs were separated into 2 groups: the control group was decanted into culture wells and the injection group was mixed with basal media and injected through a 21-gauge needle into culture wells. Microbeads were cultured for 3 weeks, and cell number and viability were measured weekly using electron and confocal microscopy. To assess effects of percutaneous injection in vivo, twenty-four male nude mice were randomly separated into 2 groups and injected with either empty microcapsules or ADSC-laden microcapsules. Mice were harvested at 1 and 3 months, and the implants were examined microscopically to assess bead and cell viability.


RESULTS
A flow rate of 5 mL/h and an electrostatic potential of 7 kV produced viable ADSC-laden microbeads of <200 μm. There were no differences in bead morphology and ADSC viability between microcapsules placed versus injected into tissue culture plates for up to 3 weeks. Microspheres implanted in a nude mouse model show durability up to 3 months with a host response around each individual sphere. ADSCs remained viable and showed signs of mitosis.


CONCLUSIONS
ADSCs can be readily cultured, encapsulated, and injected in alginate microspheres. Stem cells suspended in alginate microspheres survive in vivo and are seen to replicate in vitro.
Moyer
Hunter R
HR
Division of Plastic and Reconstructive Surgery, Emory University, Atlanta, GA 30322, USA. hmoyer@emory.edu
Kinney
Ramsey C
RC
Singh
Kimberly A
KA
Williams
Joseph K
JK
Schwartz
Zvi
Z
Boyan
Barbara D
BD
D000273:Adipose Tissue / Q000637:transplantation; D000293:Adolescent; D000464:Alginates / Q000494:pharmacology*; D000818:Animals; D002470:Cell Survival; D002648:Child; D002675:Child, Preschool; D015331:Cohort Studies; D004195:Disease Models, Animal; D004339:Drug Compounding / Q000379:methods*; D005260:Female; D005500:Follow-Up Studies; D020723:Glucuronic Acid / Q000494:pharmacology; D006603:Hexuronic Acids / Q000494:pharmacology; D006801:Humans; D020007:Implants, Experimental; D007279:Injections, Subcutaneous; D008297:Male; D051379:Mice; D008819:Mice, Nude; D008863:Microspheres; D019736:Prostheses and Implants; D033581:Stem Cell Transplantation / Q000379:methods*; D023822:Tissue Engineering / Q000379:methods*
D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid