Characterization of Human Dental Pulp Stem Cells

Characterization of the hDPSC products was examined according to recent studies [21 (link), 22 (link), 27 (link)], as described in the Supplementary Methods. As for cell morphology assay, hDPSC products in a T-75 flask (Corning) were imaged under a Primovert inverted microscope (Carl Zeiss Microscopy). The expression of MSC surface markers including human CD146, CD105, CD73, CD90, CD34, CD45, CD14, and human leukocyte antigen DR (HLA-DR), which were referred from a previous report [28 (link)], in hDPSC products was examined by flow cytometric analysis (FCM). The lineage-specific-gene expression was analyzed in the hDPSC products cultured under the specific inductive conditions into osteoblasts, chondrocytes, and adipocytes by quantitative reverse transcription polymerase chain reaction (qRT-PCR) assay. For secretome assay, hepatocyte growth factor (HGF), interleukin 6 (IL6), monocyte chemotactic protein 1 (MCP1), and sialic acid-binding immunoglobulin-type lectin 9 (SIGLEC9) in the CM of the hDPSC products were analyzed by enzyme-linked immunosorbent assay (ELISA).

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Iwanaka T., Yamaza T., Sonoda S., Yoshimaru K., Matsuura T., Yamaza H., Ohga S., Oda Y, & Taguchi T. (2020). A model study for the manufacture and validation of clinical-grade deciduous dental pulp stem cells for chronic liver fibrosis treatment. Stem Cell Research & Therapy, 11, 134.

Publication 2020

T-75 flask Primovert inverted

Adipocytes Assay Cd73 Cd90 Chondrocytes Elisa Flow cytometric Gene expression Hepatocyte growth factor Human Human leukocyte antigen dr Immunoglobulin Interleukin 6 Lectin Microscopy Monocyte chemotactic protein 1 Osteoblasts Reverse transcription polymerase chain reaction Secretome Sialic acid

Corresponding Organization :

Other organizations : Kyushu University

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Variable analysis

independent variables

Culture conditions for osteoblasts, chondrocytes, and adipocytes

dependent variables

Cell morphology
Expression of MSC surface markers (CD146, CD105, CD73, CD90, CD34, CD45, CD14, HLA-DR)
Lineage-specific gene expression in osteoblasts, chondrocytes, and adipocytes
Secretion of HGF, IL6, MCP1, and SIGLEC9

control variables

Cell culture flask (T-75 Corning)
Primovert inverted microscope (Carl Zeiss Microscopy)

controls

Previous studies [21, 22, 27, 28] used as a reference for characterization methods

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