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Adhesion, proliferation, and viability of rBMSCs in microscaffolds

YY Yanqing Yang
HH Huan He
FM Fang Miao
MY Mingwei Yu
XW Xixi Wu
YL Yuanhang Liu
JF Jie Fu
JC Junwei Chen
LM Liya Ma
XC Xiangru Chen
XP Ximing Peng
ZY Zhen You
CZ Chuchao Zhou
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For the cell adhesion evaluation, the unattached rBMSCs were carefully removed from the bottom of the plate and counted N. The cell adhesion was calculated by the equation:

5 × 105 is the total amount of cells, and N is the number of cells attached to the plate.

The cell-seeded microscaffolds were placed in 96-well plates in advance. The proliferation of rBMSCs was evaluated using MTT assays at 1, 3, and 7 day postculture. The cell viability of rBMSCs seeded in microscaffolds was evaluated by live/dead cell imaging kit staining at 1, 3, and 7 day post culture. In brief, the samples were washed with phosphate-buffered saline (PBS) and incubated in 2 μM fluorescein diacetate (FDA, Sigma, staining live cells) for 30 min and 4 μM propidium iodide (PI, Sigma, staining dead cells) for 10 min at 37 ℃. The stained samples were finally washed with PBS and observed under a confocal laser microscope (Leica, Germany).

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