Native MSC-Derived Human Chondrocytes Neuromics -Vitro Biopharma Quality Control Study Cell Culture and Viability of SC00A7 lot 006 January 2013

1. Labeled and unlabeled chondrocytes
Native MSC-Derived Human Chondrocytes Neuromics -Vitro Biopharma Quality
Control Study Cell Culture and Viability of SC00A7 lot 006 January 2013
Phase contrast image (100x) of chondrocyte and associated cells in day one (29 hours after
innoculation) of culture as described (SC00A7 Protocol). Scale bar = 50 m. Chondrocyte
is darker multicellular structure overlying attached cells, ~100 m in diameter.

2. Labeled and unlabeled chondrocytes
Phase contrast image (100x) of chondrocyte and associated cells in day two (45
hours after innoculation) of culture as described (SC00A7 Protocol). Scale bar = 50
m. Chondrocyte is darker multicellular structure overlying attached cells, ~150 m
in diameter.

3. Labeled and unlabeled chondrocytes
Phase contrast image (200x) of chondrocyte and associated cells in day five of
culture as described (SC00A7 Protocol). Scale bar = 50 m. Chondrocyte is darker
multicellular structure overlying attached cells, ~ 70 m in diameter.

4. Labeled and unlabeled chondrocytes
Cell viability of chondrocyte cultures determined by propidium iodide(PI) method in the
second day of culture. Cells were cultured as above and 1 mg/ml PI was added to yield
a final [PI] of 5 g/ml. PI positive (dead) cells are shown here as white. Viable was >/= 70%.
Scale bar is 50 m. Direct measurement of viability by the PI method at day 5 yielded
83% viability.

5. Conclusions:
• Human MSC-derived chondrocytes were maintained viable in cell culture using
Neuromics-Vitro Biopharma catalog number PC-004 Human Chondrocyte Maintenance
Medium in Greiner Bio-One TC-coated 48-well cell culture plates innoculated with 20
to 50 l/well (2,000 to 5,000 cells per well)
• Cell cultures consisted of multicellular chondrocytes together with attached cells,
that are possibly MSCs derived from chondrocytes.
• Cell viability was high throughout 5 full days of culture as determined by propidium
Iodide permeation of cells. Cellular viability of cells detached from these cultures by
Accutase treatment was 83% after five days in continuous culture.
Labeled and unlabeled chondrocytes
Culturing Protocol