Chang-Woo Lee

Multipotency and growth characteristic of periosteum-derived progenitor cells for chondrogenic, osteogenic, and adipogenic differentiation

The mesengenic multipotency of cryopreserved periosteum-derived progenitor cells (PDPCs) for chondrogenesis, osteogenesis and adipogenesis was investigated. Differentiation was verified using RT-PCR and histological analysis. For characterization, FACS analysis was performed with specific surface markers of mesenchymal stem cells (MSCs). Among PDPCs, unsorted periosteum-derived cells (PDCs) and dermal fibroblasts, the most distinct characteristics were found to be CD9, CD105, and CD166. In addition, these markers in PDPCs were continuously maintained until passage 15. We developed a rapid method for the isolation of PDPCs that can differentiate into mesodermal lineages and provide enough cells in a short period of time for allogeneic cell therapy.

Glucose-stimulated insulin secretion of various mesenchymal stem cells after insulin-producing cell differentiation

Mesenchymal stem cells (MSCs) are capable of crossing germinative layer borders and are obtainable in high numbers via in vitro cultures. Therefore, many researchers have searched for diverse sources of MSCs. Recently the generation of glucose-responsive insulin-producing cells (IPCs) from MSCs has shown immense potential for the treatment of type 1 diabetes mellitus (T1DM) due to a lack of pancreas donors. In this study, we compared the growth potency of four kinds of MSCs derived from bone marrow, Whartons jelly, adipose tissue, and the periosteum. In addition, in vitro differentiation of these MSCs into IPCs was also investigated. After 2weeks of IPCs differentiation, we compared the expression of the insulin gene and protein using RT-qPCR and immunofluorescence staining. Only IPCs derived from periosteum-derived progenitor cells (PDPCs) showed a response to glucose concentration. Glucose stimulated insulin secretion was conclusive evidence of the potential functionality of IPCs. Therefore, PDPCs are a promising alternative stem cell source for IPCs differentiation.

Isolation of human periosteum-derived progenitor cells using immunophenotypes for chondrogenesis

Periosteum-derived progenitor cells (PDPCs) were isolated by characteristic surface markers. Reproducibility of immunophenotypes of the PDPCs was characterized by flow cytometric analysis using fluorescence-activated cell sorter (FACS). SH2+, SH3+, SH4+, CD9+, CD90+ and CD105+ were important eternal characteristic cell surface markers for the PDPCs. The characterized PDPCs maintained their chondrogenic potential in pellet cultures until the 15th passage from primary cell culture.

Chondrogenesis of human periosteum-derived progenitor cells in atelocollagen.

Periosteum-derived progenitor cells (PDPCs) could be differentiated into cartilage using atelocollagen as a carrier and in the presence of transforming growth factor-β3 (TGF-β3). Chondrogenesis was verified by RT-PCR and Western blotting. Expression of the type II collagen mRNA was found from the differentiated PDPCs in atelocollagen 3xa0weeks after chondrogenic induction. The chondrogenic potential of the PDPCs was also verified by histochemical staining for type II collagen protein. Increased production of glycosaminoglycan shows that the PDPCs in atelocollagen could differentiate into chondrocytes under a chondrogenic environment. PDPCs can therefore be used as a cell source for cell-based therapies targeted toward the articular cartilage of the knee.

Chondrogenic properties of human periosteum-derived progenitor cells (PDPCs) embedded in a thermoreversible gelation polymer (TGP)

Periosteum-derived progenitor cells (PDPCs) were isolated using a fluorescence-activated cell sorter and their chondrogenic potential in biomaterials was investigated for the treatment of defective articular cartilage as a cell therapy. The chondrogenesis of PDPCs was conducted in a thermoreversible gelation polymer (TGP), which is a block copolymer composed of temperature-responsive polymer blocks such as poly(N-isopropylacrylamide) and of hydrophilic polymer blocks such as polyethylene oxide, and a defined medium that contained transforming growth factor-β3 (TGF-β3). The PDPCs exhibited chondrogenic potential when cultured in TGP. As the PDPCs-TGP is an acceptable biocompatible complex appropriate for injection into humans, this product might be readily applied to minimize invasion in a defected knee.

Effects of silkworm hemolymph and cartilage-specific extracellular matrices on chondrocytes and periosteum-derived progenitor cells

In order to enhance the repair of defects in articular cartilage via cell therapy with autologous chondrocytes, as well as with periosteum-derived progenitor cells (PDPCs), silkworm hemolymph (SH) and a variety of cartilage-specific extracellular matrices (ECMs) including type II collagen, proline, chondroitin 4-sulfate, and chondroitin 6-sulfate were assessed with regard to their efficacy as media supplements. SH, a known anti-apoptotic agent, was found to enhance cell growth, as was shown by the results of a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay. According to the results of reverse transcriptase polymerase chain reaction (RT-PCR) analyses, the cartilage-specific ECMs were found to stimulate the expression of hyaline cartilage-specific genes, most notably type II collagen and Sox9, in monolayer cultures of PDPCs.

Pancreatic islet-like clusters from periosteum-derived progenitor cells

Recent studies comparing the insulin-producing cell (IPC) differentiation capacity of mesenchymal stem cells (MSCs) derived from four different sources (bone marrow, Wharton’s jelly, adipose tissue, and the periosteum) demonstrated that IPC differentiation of periosteum-derived progenitor cells (PDPCs) progressed faster than any other MSCs within 7 days, indicating that PDPCsare most suited to IPC differentiation. Here, two different cell culture methods, adhesion and cluster culture, were assessed for their ability to support in vitro IPC differentiation. The induction of IPC differentiation was confirmed by RTqPCR analysis of insulin gene expression levels and immunofluorescence analysis of insulin protein. An enzyme-linked immunosorbent assay was used to quantify secreted insulin. PDPC-derived IPCs from cluster cultures demonstrated a significantly increased expression of insulin and an enhanced secretion of insulin of insulin protein in response to glucose compared to IPCs derived from adhesion cultures. Thus, pancreatic islet-like cluster cultures appear to provide the optimal conditions such as cluster culture for IPC differentiation of PDPCs.