title: Bone marrow and adipose stem cells can be tracked with PKH26 until post staining passage 6 in in vitro and in vivo.
abstract: Tracking of transplanted cells has become an important procedure in cell therapy. We studied the in vitro dye retention, survival and in vivo tracking of stem cells with PKH26 dye. Sheep BMSCs and ADSCs were labeled with 2, 4 and 8 μmol of PKH26 and monitored for six passages. Labeled BMSCs and ADSCs acquired mean cumulative population doubling of 12.7±0.4 and 14.6±0.5; unlabeled samples had 13.8±0.5 and 15.4±0.6 respectively. Upon staining with 2, 4 and 8 μmol PKH26, BMSCs had retentions of 40.0±5.8, 60.0±2.9 and 95.0±2.9%, while ADSCs had 92.0±1.2, 95.0±1.2 and 98.0±1.2%. ADSCs retentions were significantly higher at 2 and 4 μmol. On dye retention comparison at 8 μmol and 4 μmol for BMSCs and ADSCs; ADSCs were significantly higher at passages 2 and 3. The viability of BMSCs reduced from 94.0±1.2% to 90.0±0.6% and ADSCs from 94.0±1.2% to 52.0±1.2% (p<0.05) after 24h. BMSCs had significant up regulation of the cartilage genes for both the labeled and the unlabeled samples compared to ADSCs (p<0.05). PKH26 fluorescence was detected on the resected portions of the regenerated neo-cartilage. The recommended concentration of PKH26 for ADSCs is 2 μmol and BMSCs is 8 μmol, and they can be tracked up to 49 days.
authors.affiliation: Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latif, Bandar Tun Razak, Cheras 56000, Kuala Lumpur, Malaysia.
authors.forename: C C
authors.initials: CC
authors.lastname: Ude
authors.forename: B S
authors.initials: BS
authors.lastname: Shamsul
authors.forename: M H
authors.initials: MH
authors.lastname: Ng
authors.forename: H C
authors.initials: HC
authors.lastname: Chen
authors.forename: M Y
authors.initials: MY
authors.lastname: Norhamdan
authors.forename: B S
authors.initials: BS
authors.lastname: Aminuddin
authors.forename: B H I
authors.initials: BH
authors.lastname: Ruszymah
mesh_terms: D000273:Adipose Tissue / Q000737:chemistry / Q000166:cytology*; D000818:Animals; D001854:Bone Marrow Cells / Q000737:chemistry*; D002454:Cell Differentiation; D002470:Cell Survival; D058948:Cell Tracking / Q000379:methods*; D002478:Cells, Cultured; D020219:Chondrogenesis; D003470:Culture Media; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry; D005786:Gene Expression Regulation; D009930:Organic Chemicals / Q000737:chemistry*; D012756:Sheep; D013194:Staining and Labeling / Q000379:methods*; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000737:chemistry*
chemical_list: D003470:Culture Media; D005456:Fluorescent Dyes; D009930:Organic Chemicals; C070080:PKH 26