Quantifying Organ-Specific Cell Migration

Standard curves for the evaluation of the migratory ability of intraperitoneally injected cells were generated by administering serial dilutions of hAT-MSCs to mouse tissues, as described previously^{14 (link), 16 (link)}. Briefly, 2 × 10, 2 × 10², 2 × 10³, 2 × 10⁴, or 2 × 10⁵ hAT-MSCs were added to the whole mouse organs prior to homogenization. After the total RNA was extracted from the samples (Easy-BLUE Total RNA Extraction kit; Intron Biotechnology), cDNA was synthesised (LaboPass M-MuLV Reverse Transcriptase; Cosmo Genetech) using 1 μg of RNA. qRT-PCR using human-specific GAPDH primers (forward primer, 5′-TGC TTT TAA CTC TGG TAA AGT GGA TA-3′; reverse primer, 5′-GTG GAA TCA TAT TGG AAC ATG TAA AC-3′) was performed in order to generate the standard curve. The curve was corrected by performing parallel qRT-PCR with primers used to amplify both human and mouse GAPDH (forward primer, 5′-CAG CGA CAC CCA CTC CTC CAC CTT-3′; reverse primer, 5′-CAT GAG GTC CAC CAC CCT GTT GCT-3′).