Differentiation of isolated and characterized human dental pulp stem cells and stem cells from human exfoliated deciduous teeth: An in vitro study

Abstract

Aims and objectives:Isolation, characterization and differentiation of dental pulp stem cells (DPSCs) and stem cells from exfoliated human deciduous teeth (SHED).Methods:The pulp tissue was digested in collagenase and cultured in DMEM Dulbecco's Modified Eagle's Media). The stem cells were identified and isolated. Surface characterization of cells was done with flow cytometer using surface markers. An immuno cytochemistry analysis was done. Differentiation potential was analyzed using various differentiation markers.Results:Flow cytometry analyses for various CD markers showed similar results for both DPSCs and SHED. The cells showed positive expression for pluripotent, ectodermal and mesodermal markers. Cells differentiated into osteoblasts and adipocytes.Conclusion:The study demonstrated that stem cells existed in deciduous and permanent pulp tissue. The stem cells present in pulp tissue can be isolated, cultivated and expanded in vitro. Both DPSCs and SHED show almost a similar expression pattern profile for variety of antigens tested.

Epidermal growth factor enhances osteogenic differentiation of dental pulp stem cells in vitro

“…It is noteworthy that after 1 week in osteogenic conditions, the DPSCs changed their colony-cell distribution; moreover, a greater cell adherence can be observed. As a general consensus, some surface markers are included within the minimum criteria for defining MSCs [ 48 ]; however, others markers have been associated with MSC lineage, such as CD146 and CD10, both expressed on DPSCs [ 49 , 50 ] but their biological implication to the MSC lineage remains poorly known. Furthermore, in vitro EGF treatment was enough to reduce the expression of both cell markers, confirming an osteogenic role by EGF on DPSCs.…”

Insight into the Role of Dental Pulp Stem Cells in Regenerative Therapy

“…DPSCs express MSC-related markers, such as CD13, CD44, CD73, CD90, CD146, CD166, and STRO-1 [ 9 , 16 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 ], while they do not express monocytic and hematopoietic lineage markers, such as CD14, CD19, CD34, CD45, and HLA-DR surface molecules [ 9 , 16 , 55 , 56 , 63 ]. SHEDs have been reported to show an expression pattern similar to DPSCs, such as CD13, CD44, CD73, CD90, CD146, CD166, and STRO-1 [ 10 , 16 , 17 , 56 , 57 , 61 , 62 ], but not CD34, CD45, or HLA-DR [ 16 , 17 , 56 , 57 ]. The absence of these monocytic and hematopoietic lineage markers is a defining feature of mesenchymal cells.…”

Effects of a discoloration-resistant calcium aluminosilicate cement on the viability and proliferation of undifferentiated human dental pulp stem cells

“…The dental pulps were minced and digested in a solution containing 3 mg/mL type I collagenase and 4 mg/mL dispase (Gibco BRL, Gaithersburg, MD, USA) at 37 °C for 2 h. Single-cell suspensions were obtained by passing the cells through a 70-mm strainer (BD Falcon, Franklin Lakes, NJ, USA) and cultured in growth medium (α-modified Eagle medium (Gibco) supplemented with 10% fetal bovine serum (Gibco), 100 units/mL penicillin and 100 mg/mL streptomycin) in 5% CO 2 at 37 °C. To identify hDPSCs, the cultured cells were incubated with fluorescent dye-conjugated monoclonal antibodies for different Cluster of Differentiation (CD) cell-surface molecular markers, including anti-CD29, anti-CD34, anti-CD44, anti-CD45, anti-CD90 and anti-CD105 (EMD Millipore Corp., Billerica, MA, USA), and sorted using a flow cytometer (Elite ESP, Beckman Coulter, Fullerton, CA, USA) 32 33 . To confirm the specificity of primary antibody binding nonspecific mouse IgM isotype control (lambda monoclonal MOPC-104E, Abcam, Cambridge, MA, USA) which matches the primary antibody’s host species was substituted for the primary antibody.…”

“…Thus, the objective of the present study was to examine the effects of the experimental discoloration-resistant calcium aluminosilicate cement on the viability and proliferation of human dental pulp stem cells (hDPSCs) prior to their differentiation. Although hDPSCs are multipotent and have the capacity to differentiate into chondrogenic, adipogenic and osteogenic cells, the well-being of the original stem cells is a prerequisite for these events to occur 32 33 . The null hypothesis tested was that there is no difference in the various facets of cytotoxicity induced by the experimental calcium aluminosilicate cement and a calcium silicate endodontic cement when these set cements are placed in close proximity with undifferentiated hDPSCs.…”