Human BM-MSCs used for in vitro viability experiments were obtained
from Lonza, Walkersville, MD. Cells were initially isolated from adult bone
marrow of bilateral iliac crests of normal, non-diabetic volunteers and
cryopreserved after passage 2. Passage 9 cells were encapsulated in
RLP-AM/HA-SH hydrogels at a density of 1 × 106cells/mL.
In vivo experiments were completed using BM-MSCs with demonstrated
success in rabbit vocal fold scar [52 (link)]. Human BM-MSCs were obtained from Waisman Biomanufacturing,
Madison, WI. As previously described [52 (link), 53 (link)], cells were
derived from the iliac crest of a healthy 22-year-old female and expanded
and tested using procedures that met Current Good Manufacturing Practices
(cGMPs) for human clinical trials. Cells were suspended in cold freezing
medium containing PlasmaLyte, 10% rabbit serum albumin, and 2.5% dimethyl
sulfoxide (DMSO) at 2 × 107 cells/mL and cryopreserved in
liquid nitrogen. Passage 6 cells were used in this study. Cells were thawed
just before injection, when they were gently mixed with RLP-AM/HA-SH gel via
pipetting at a final concentration of 3 × 105 cells in 100
μL injection volume.
from Lonza, Walkersville, MD. Cells were initially isolated from adult bone
marrow of bilateral iliac crests of normal, non-diabetic volunteers and
cryopreserved after passage 2. Passage 9 cells were encapsulated in
RLP-AM/HA-SH hydrogels at a density of 1 × 106cells/mL.
In vivo experiments were completed using BM-MSCs with demonstrated
success in rabbit vocal fold scar [52 (link)]. Human BM-MSCs were obtained from Waisman Biomanufacturing,
Madison, WI. As previously described [52 (link), 53 (link)], cells were
derived from the iliac crest of a healthy 22-year-old female and expanded
and tested using procedures that met Current Good Manufacturing Practices
(cGMPs) for human clinical trials. Cells were suspended in cold freezing
medium containing PlasmaLyte, 10% rabbit serum albumin, and 2.5% dimethyl
sulfoxide (DMSO) at 2 × 107 cells/mL and cryopreserved in
liquid nitrogen. Passage 6 cells were used in this study. Cells were thawed
just before injection, when they were gently mixed with RLP-AM/HA-SH gel via
pipetting at a final concentration of 3 × 105 cells in 100
μL injection volume.
