MYOBLAST SEEDING IN A COLLAGEN MATRIX EVALUATED IN-VITRO

Collagens may be used as biomaterials for soft tissue reconstruction, e.g., the abdominal wall. We previously developed a biocompatible derm al sheep collagen (DSC), which in an abdominal wall reconstruction mod el showed controlled biodegradation and functioned as a matrix for ing rowth of fibroblasts but not of muscle. It was hypothesized that regen eration of muscle via DSC may be possible by seeding of muscle cells. Using a syringe, mouse C2C12 myoblasts were seeded in DSC disks and in cubated in methylcellulose-based growth medium, changed at 24 h into d ifferentiation medium. An estimated 85% of the cells were well distrib uted, especially in the top half of the DSC disks. Some 15% of the cel ls ended up on top. At 4 h, all cells showed a spherical morphology, s ometimes with clear adhesion plaques. At 24 h, cells on the top starte d to form a ''capsule'' with well-spread cells. Underneath the capsule , of the remaining 85% of the cells, approximately 30% showed adhesion and spreading on/in between collagen bundles. At day 3 after the addi tion of differentiation medium, the spread cells showed first indicati ons of myotube formation. At day 7, myotube formation had proceeded, w hile extracellular matrix, i.e., collagen and elastin, had been deposi ted. This study shows that myoblast seeding into DSC is feasible, resu lting in a reasonable cell distribution and survival of 45% of the cel ls. The surviving cells are able to differentiate into myotubes and fo rm an extracellular matrix. (C) 1996 John Wiley & Sons, Inc.