Human Dental Pulp Stem Cells: The Culture Optimization for Increased Growth
,
Abstract
Introduction: Dental pulp-derived mesenchymal stem cells (MSCs) have emerged as a promising tool for use in regenerative medicine. The in vitro growth kinetic and culture requirement of the cells derived from human dental pulp, which is the subject of this present study, is poorly described.
Materials and Methods: Stem cells were derived from human third molar and then characterized. The in vitro growth kinetics of the cells was examined by colonogenic assay and a determination of the population doubling number (PDN). Finally, the culture conditions were optimized for pulp stem cell maximum proliferation.
Results: Propagated dental pulp cells tended to differentiate into odontoblast, osteoblast, adipose and cartilage cells. Typically surface antigens were expressed as mesenchymal stem cells. The cells tended to be very proliferative with a PDN value of about 11. The colonogenic efficiency was about 60% and an average colony size was about 10.75±1.58 mm2. The best culture condition for enhanced proliferation was achieved when the cells were seeded at 100 cells/cm2 in a the presence of 20% FBS in a medium (P<0.05).
Conclusion: Taken together, the optimal culture conditions for human dental pulp-derived MSCs were determined. This information is helpful with respect to cell in vitro propagation which is greatly needed prior to their transplantation.
Baksh D, Song L, Tuan RS. Adult Mesenchymal Stem Cells: Characterization, Differentiation and Application in Cell Therapy. Mol Med, 2004; 8: 301-136.
Horwitz EM, Gordon PL, Koo WK, et al. Isolated Allogenic Bone Marrow- drived Mesenchymal Cells Engraft and Stimulate Growth in Children with Osteogenesis Imperfecta: Implications for Cell Therapy of Bone. Proc Natl Acad Sci, USA, 2002; 99: 8932-8937.
Koe ON, Gerson SL, Cooper BW, et al. Rapid Hematopoietic Recovery after Confusion of Autologous - Blood Stem Cells in Advanced Breast Cancer Patients Receiving High Dose Chemotherapy. J Clin Oncol, 2000; 18: 307-316.
Quarto R, Mastrogiacomo M, Cancedda R, et al. Repair of Large Bone Defect with the Use of Autogenic Bone Marrow Stromal Cell. N Engl J Med, 2001; 344: 385-386.
Barry FP. Mesenchymal Stem Cell Therapy in JointDisease. Novartis found symp, 2003; 249: 86-96.
Sugaya K. Potential Use of Stem Cells in Neuro- Replacement Therapies for Neurodegenerative Diseases. Int Rev Cytol, 2003; 228: 1-30.
Chapel A, Bertho JM, Bensidhoum A, et al. Mesenchymal Stem Cells Home to Injured Tissues when co-Infused with Hematopoietics Cells to Treat a Radiation- Induced Multi- Organ Failure Syndrome. J Gene Med, 2003; 5: 1028-1038.
Pittenger MF, Mackay AM, Beck SC, et al. Multilineage Potential of Adult Human Mesenchymal Stem Cells. Science, 1999; 284: 143-147.
Baxter MA, Wynn RF, Jowitt SN, et al. Study of Telomere Length Reveals Rapid Aging of Human Marrow Stromal Cells Following in Vitro Expansion. Stem Cells, 2004; 22: 675-82.
Nishida S, Endo N, Yamagiwa H, et al. Number of Osteoprogenitor Cells in Human Bone Marrow Markedly Decreases after Skeletal Maturation. J Bone Miner Metab, 1999; 17: 171-177.
Noth U, Osyczka AM, Tuli R, et al. Multilineage Mesenchymal Differentiation of Human Trabecular Bone- Derived Cells. J Ortho Res, 2002; 20: 1060-1069.
Wickham MQ, Erickson GR, Gimble JM, et al. Multipotent Stromal Cells Derived from Infrapatellar Fat Pad of Knee. Clin Ortho, 2003; 196-212.
Jankowsk RJ, Deasy BM, Huard J. Muscle-Derived Stem Cells. Gene therapy, 2002; 9: 642-647.
Zvaifler NJ, Marinova-Mutafchieva L, Adams G, et al. Mesenchyamal Precursor Cells in the Blood of Normal Individuals. Artheritis Res, 2000; 2: 477-488.
You Q, Tong X, Guan Y, et al. B J Int Med Res, 2009; 37: 105-112.
Erices A, Conget P, Miguell JJ. Mesenchymal Progenitor Cells in Human Umbilical Cord Blood. Br J Haematol, 2000; 109: 235-242.
Eslaminejad MB, Taghiyar L. Mesenchymal Stem Cell Purification from the Articular Cartilage Cell Culture. Iran J Basic Med. Sci, 2008; 3: 146-153.
Gronthos S, Mankani M, Brahim J, et al. Postnatal Human Dental Pulp Stem Cells (DPSCs) in Vitro and in Vivo. Proc Natl Acad Sci, U S A, 2000; 97: 13625-13630.
Laino G, Carinci F, Graziano A, et al. In Vitro Bone Production Using Stem Cells Derived from Human Dental Pulp. J Craniofac Surg, 2006; 17: 511-515.
d’Aquino R, Graziano A, Sampaolesi M, et al. Human Postnatal Pulp Cells co-Differentiate into Osteoblast and Endotheliocytes: A Pivotal Synergy Leading to Adult Bone Tissue Formation. Cell Death Differ, 14: 1162-1171.
Zhang W, Walboomers XF, Shi S, et al. Multilineage Differentiation Potential of Stem Cells Derived from Human Dental Pulp after Cryopreservation. Tissue Eng, 2006; 12: 2813-2823.
Huang GT, Gronthos S, Shi S. Mesenchymal Stem Cells Derived from Dental Tissues vs those from other Sources: Their Biology and Role in Regenerative Medicine. J Dent Res, 2009; 88: 792-806.
Buurma B, Gu K, Rutherford RB. Transplantation of Human Pulpal and Gingival Fibroblasts Attached to Synthetic Scaffolds. Eur J Oral Sci, 1999; 107: 282-289.
Gu K, Smoke RH, Rutherford RB. Expression of Genes for Bone Morphogenetic Proteins and Receptors in Human Dental Pulp. Arch Oral Biol, 1996; 41: 919-923.
Myers WC, Fountain SB. Dental Pulp Regeneration Aided by Blood and Blood Substitutes after Experimentally Induced Periapical Infection. Oral Surg Oral Med Oral Pathol, 1974; 37: 441-450.
Ostby BN. The Role of the Blood Clot in Endodontic Therapy. An Experimental Histologic Study. Acta Odontol Scand, 1961; 19: 324-353.
Mooney DJ, Powell C, Piana J, Rutherford B. Engineering Dental Pulp-like Tissue in Vitro. Biotechnol Prog, 1996; 12: 865-868.
Eslaminejad MB, Nikmahzar A, Thagiyar L, et al. Murine Mesenchymal Stem Cells Isolated by Low Density Primary Culture System. Dev Growth Diff, 2006; 48: 361-370.
Tonomura A, Sumita Y, Ando Y, et al. Differential Inducibility of Human and Porcine Dental Pulp-derived Cells into Odontoblasts. Connect Tissue Res, 2007; 48: 229-238.
Piera-Velazquez S, Jimenez SA, Stokes D. Increased Life Span of Human Osteoarthritic Chondrocytes by Exogenous Expression of Telomerase. Arthritis Rheum, 2002; 46: 683- 693.
Dominici M, Le Blanc K, Mueller I, et al. Minimal Criteria for Defining Multipotent Mesenchymal Stromal Cells. The International Societry for Cellular Therapy Position Statement. Cytotherapy, 2006; 4: 315-317.
Spees JL, Gregory CA, Singh H, et al. Internalized Antigens must be Removed to Prepare Hypoimmunogenic Mesenchymal Stem Cells for Cell and Gene Therapy. Mol Ther, 2004; 95: 747-756.
Pittenger MF, Mackay AM, Beck SC, et al. Multilineage Potential of Adult Human Mesenchymal Stem Cells. Science, 1999; 284: 143-147.
Bartmann C, Rohde E, Schallmoser K, et al. Relationship between Donor Age and the Replicative Lifespan of Human Cells in Culture: A Reevaluation. Proc Natl Acad Sci, USA, 1998; 95: 10614-9.
| Files | ||
| Issue | Vol 3, No 4 (2009) | |
| Section | Articles | |
| Keywords | ||
| Dental Pulp Human Third Molar Mesenchymal Stem Cells Odontoblast Proliferation Skeletal Cell | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
