Enzymatically fabricated and degradable microcapsules for production of multicellular spheroids with well-defined diameters of less than 150 microm.
Microcapsules with a single, spherical hollow core less than 150 microm in diameter were developed to obtain multicellular spheroids with well-defined sizes of less than 150 microm in diameter. An aqueous solution of phenolic hydroxyl derivative of carboxymethylcellulose (CMC-Ph) containing human hepatoma cell line (HepG2) cells and horse radish peroxidase (HRP) was injected into a coflowing stream of liquid paraffin, containing H(2)O(2), resulting in cell-enclosing CMC-Ph microparticles, 135 microm in diameter, via a peroxidase-catalyzed crosslinking reaction. The CMC-Ph microparticles were then coated with a phenolic hydroxyl derivative of alginate (Alg-Ph) gel membrane several dozen micrometers in thickness, crosslinked via the same enzymatic reaction process, followed by further crosslinking between the carboxyl groups of alginate by Sr(2+). A hollow core structure was achieved by immersing the resultant microcapsules in a medium containing cellulase, which degrades the enclosed CMC-Ph microparticles. The HepG2 cells in the microcapsules then grew and completely filled the hollow core. Multicellular spheroids the same size as the CMC-Ph microparticles, with living cells at their outer surface, were collected within 1 min by soaking them in a medium containing alginate lyase to degrade the Alg-Ph gel microcapsule membrane.
Sakai
Shinji
S
Department of Chemical Engineering, Faculty of Engineering, Kyushu University, Nishi-ku, Fukuoka, Japan. sakai@chem-eng.kyushu-u.ac.jp
Ito
Sho
S
Ogushi
Yuko
Y
Hashimoto
Ichiro
I
Hosoda
Natsuko
N
Sawae
Yoshinori
Y
Kawakami
Koei
K
D000464:Alginates / Q000737:chemistry; D001672:Biocompatible Materials / Q000737:chemistry*; D001673:Biodegradation, Environmental; D002214:Capsules / Q000737:chemistry*; D002266:Carboxymethylcellulose Sodium / Q000737:chemistry; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D002470:Cell Survival; D002480:Cellulase / Q000737:chemistry; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D013324:Strontium / Q000737:chemistry; D013997:Time Factors; D014783:Viscosity
D000464:Alginates; D001672:Biocompatible Materials; D002214:Capsules; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D006861:Hydrogen Peroxide; D002480:Cellulase; D002266:Carboxymethylcellulose Sodium; D013324:Strontium