| | MSC type | First found time | Source | Extraction approach | Culture medium | Marker | Reference |
| | DPSCs | 2002 | Pulp of human third molars | DPSC-ED | (1) Tooth surfaces were cleaned and cut around the cementum-enamel junction by using sterilized dental fissure burs to reveal the pulp chamber
(2) Rinse with culture medium and cut dental pulp tissue into small pieces of 1-2 mm3
(3) Pulp fragments were digested in a solution of 3 mg/ml collagenase type I and 4 mg/ml dispase II for 1 h at 37°C
(4) After digestion, it was centrifuged at a speed of 300 g, and the resuspended tissue fragments were passed through 70 μM cell filter
(5) The cells were cultured in 6-well plates in standard DPSC medium. All cell cultures were incubated at 37°C in 5% CO2 | Alpha modification of Eagle’s medium supplemented with 10% FBS+100 μM L-ascorbic acid 2-phosphate+2 mM L-glutamine+100 U/ml penicillin+100 μg/ml streptomycin | Positive: CD29, CD44, CD49f, CD73, CD81, CD90, CD105, and CD146
Negative: CD14, CD19, CD34, CD45, Stro-1, and HLA-DR | [8, 14–19] | | DPSC-OG | (1) After disinfecting the tooth surface, mechanically break the tooth and gently separate the dental pulp with tweezers (there is no need to drill, because it may adversely affect the viability of dental pulp stem cells)
(2) Rinse with culture medium and cut dental pulp tissue into small pieces of 1-2 mm3
(3) Pulp pieces were cultured in 6-well plates in standard DPSC medium. All cell cultures were incubated at 37°C in 5% CO2 |
| | DFSCs | 2005 | Normal human impacted third molars | (1) Normal human impacted third molars were surgically removed and collected. Attached dental follicles were separated from the mineralized tooth
(2) The surfaces of the follicle tissues were cleaned and minced by using a sterilized scalpel (dental papilla tissue was discarded)
(3) Tissues were digested in a solution of 3 mg/ml collagenase type I and 4 mg/ml dispase II for 1 h at 37°C
(4) Minced and digested tissues of dental follicle explants were seeded into 60 mm plates or T25 flasks in media at 37°C in 5% CO2 in a humidified atmosphere | Alpha modification of Eagle’s medium supplemented with 10% FBS+100 μM L-ascorbic acid 2-phosphate+2 mM L-glutamine+100 units/ml penicillin+100 μg/ml streptomycin | Positive: CD105, CD29, CD90, and CD73
Negative: CD45, CD19, HLA-DR, CD14, and CD34 | [9, 20–23] |
| | SCAPs | 2006 | Normal human impacted third molars | (1) Root apical papilla was gently separated from the surface of the root
(2) Tissues were minced and digested in a solution of 3 mg/ml collagenase type I and 4 mg/ml dispase for 30 minutes at 37°C
(3) Single cell suspensions of SCAP were obtained by passing through a 70 μM strainer
(4) Minced and digested tissues of dental follicle explants were seeded into 60 mm plates or T25 flasks in media at 37°C in 5% CO2 in a humidified atmosphere | Alpha modification of Eagle’s medium supplemented with 15% FBS+100 μM L-ascorbic acid 2-phosphate+2 mM L-glutamine+100 U/ml penicillin+100 μg/ml streptomycin | Positive: STRO-1, CD24, CD29, CD73, CD90, CD105, CD106, CD146, CD166, and ALP
Negative: CD34, CD45, CD18, and CD150 | [10] | | Minipigs: canine | (1) The canines of Wuzhishan minipigs were extracted, and the root apical papilla was gently separated from the surface of the root
(2) Apical papilla was minced and digested in a solution of 3 mg/ml collagenase type I and 4 mg/ml dispase for 30 minutes at 37°C
(3) Then passed through a 70 μm strainer to obtain a single cell suspension and seeded into 25 cm2 culture flasks containing an basic medium | Positive:STRO-1, CD146, CD24 | [24] |
| | SHEDs | 2003 | Intact caries free primary teeth | (1) The pulp was separated from a remnant crown under strict aseptic conditions
(2) The pulp was minced by using a sterilized scalpel
(3) Tissues were digested in a solution of 3 mg/ml collagenase type I and 4 mg/ml dispase for 30 minutes at 37°C
(4) Then seeded into 25 cm2 culture flasks containing an basic medium | Dulbecco’s modified Eagle’s medium supplemented with 10% FBS+100 U/ml penicillin+100 μg/ml streptomycin | Positive: STRO-1, STRO-3, CD13, CD29, CD44, CD73, CD90, CD105, CD106, CD166, CD271, CD146, ALP, MEPE, bFGF, and endostatin
Negative: CD3, CD8, CD11b (or CD14), CD15, CD19 (or CD79α), CD33, CD34, CD45, CD71, CD117, and HLA-DR | [14, 25–27] |
| | PDLSCs | 2004 | Normal impacted third molars | (1) PDL was gently separated from the surface of the root
(2) Tissues was minced and digested in a solution of 3 mg/ml collagenase type I and 4 mg/ml dispase for 1 hour at 37°C
(3) Single cell suspensions of PDLSCs were obtained by passing through a 70 μM strainer
(4) Minced and digested tissues were seeded into 60 mm plates or T25 flasks in media at 37°C in 5% CO2 in a humidified atmosphere | Alpha modification of Eagle’s medium supplemented with 10% FBS+100 μM L-ascorbic acid 2-phosphate+2 mM L-glutamine+100 units/ml penicillin+100 μg/ml streptomycin | Positive: CD105, CD29, CD90, and CD73
Negative: CD45, CD19, HLA-DR, CD14, and CD34 | [11, 22, 28, 29] |
| | GMSCs | 2009 | Attached keratinized gingival tissues | (1) The tissues were deepithelialized and minced into 1–2 mm2 fragments
(2) The minced tissues were digested in 2 mg/ml collagenase and 1 mg/ml dispase for 30 min
(3) After discarding the first digested cell suspension, the tissues were digested in the same solution for 90 min at 37°C
(4) Single cell suspensions of GMSCs were obtained by passing through a 70 μM strainer
(5) Minced and digested tissues were seeded into 60 mm plates or T25 flasks in media at 37°C in 5% CO2 in a humidified atmosphere | Alpha modification of Eagle’s medium containing 15% FBS+100 U/ml penicillin+100 μg/ml streptomycin +200 mM L-glutamine +10 mM ascorbic acid 2-hosphate | Positive: CD44, CD73, CD90, CD105, SSEA-4, STRO-1, CD146, CD166, and CD271
Negative: CD14, CD45, CD34, and CD19 | [30] |
| | OMSCs | 2005 | Human upper middle turbinates | (1) All biopsies were collected on ice in Hanks’ balanced salt solution containing penicillin (100 U/ml), streptomycin (100 mg/ml), and Fungizone (amphotericin B, 1.25 mg/ml)
(2) After being minced with a scalpel blade, the tissue was digested using 1.33% collagenase for 20 min
(3) Then, the tissues were incubation with DNAse to reduce cell clumping (0.04 mg/ml bovine pancreas DNAse, 3.0 mg/ml bovine serum albumin-fraction A in L15)
(4) Cells were mechanically dissociated by pipetting and then triturating through a 23G needle and centrifuged at 1200 rpm for 5 min, and the pellet resuspended in low-glucose Dulbecco’s modified Eagle’s medium | Low-glucose Dulbecco’s modified Eagle’s medium supplemented with 10% FBS+100 U/ml penicillin+100 μg/ml streptomycin | Positive: CD90, CD54, CD105, CD73, nestin, CD166, and p75NTR
Negative: STRO-1 | [13] |
| | JMSCs | 2006 | Human undergoing orthognathic surgery | The resected bone mass was cut into small fragments (<1 mm3) and cultured in T25 flasks with α-MEM containing 10% FBS in a 37°C humidified incubator with a 5% CO2 atmosphere | Alpha modification of Eagle’s medium containing 10% FBS+100 U/ml penicillin+100 μg/ml streptomycin | Positive: CD73, CD90, and CD105
Negative: CD34, CD11b, CD19, CD45, and HLA-DR | [31–34] |
|
|
