Cryopreservation Method for the Effective Collection of Dental Pulp Stem Cells.
Dental pulp stem cells (DPSCs) are an attractive cell source for use in cell-based therapy, regenerative medicine, and tissue engineering because DPSCs have a high cell proliferation ability and multidifferentiation capacity. However, several problems are associated with the collection and preservation of DPSCs for use in future cell-based therapy. In particular, the isolation of DPSCs for cryopreservation is time consuming and expensive. In this study, we developed a novel cryopreservation method (NCM) for dental pulp tissues to isolate suitable DPSCs after thawing cryopreserved tissue. Using the NCM, dental pulp tissues were cultured on adhesion culture dishes for 5 days and then cryopreserved. After thawing, the cryopreserved dental pulp tissue fragments exhibited cell migration. We evaluated each property of DPSCs isolated using the NCM (DPSCs-NCM) and the explant method alone without cryopreservation (DPSCs-C). DPSCs-NCM had the same proliferation capacity as DPSCs-C. Flow cytometry (FACS) analysis indicated that both DPSCs-NCM and DPSCs-C were positive for mesenchymal stem cell markers at the same level but negative for hematopoietic cell markers. Moreover, both DPSCs-NCM and DPSCs-C could differentiate into osteogenic, chondrogenic, and adipogenic cells during culture in each induction medium. These results suggest that DPSCs-NCM may be mesenchymal stem cells. Therefore, our novel method might facilitate the less expensive cryopreservation of DPSCs, thereby providing suitable DPSCs for use in patients in future cell-based therapies.
1 Department of Oral Medicine and Stomatology, Tsurumi University School of Dental Medicine , Yokohama, Japan .
Yusuke
Y
Takebe
1 Department of Oral Medicine and Stomatology, Tsurumi University School of Dental Medicine , Yokohama, Japan .
Seiko
S
Tatehara
1 Department of Oral Medicine and Stomatology, Tsurumi University School of Dental Medicine , Yokohama, Japan .
Tatsuhiro
T
Fukushima
1 Department of Oral Medicine and Stomatology, Tsurumi University School of Dental Medicine , Yokohama, Japan .
Reiko
R
Tokuyama-Toda
2 Division of Pathology, Department of Oral Diagnostic Sciences, School of Dentistry, Showa University , Tokyo, Japan .
Rika
R
Yasuhara
2 Division of Pathology, Department of Oral Diagnostic Sciences, School of Dentistry, Showa University , Tokyo, Japan .
Kenji
K
Mishima
2 Division of Pathology, Department of Oral Diagnostic Sciences, School of Dentistry, Showa University , Tokyo, Japan .
Kazuhito
K
Satomura
D050156:Adipogenesis / Q000502:physiology; D000293:Adolescent; D000328:Adult; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000502:physiology; D015925:Cryopreservation / Q000379:methods*; D003782:Dental Pulp / Q000166:cytology*; D006801:Humans; D010012:Osteogenesis / Q000502:physiology; D013234:Stem Cells / Q000166:cytology*; D023822:Tissue Engineering; D055815:Young Adult
cryopreservation; dental pulp stem cell; dental pulp tissue; explant method