Human Chorion-derived Stem Cells Research Articles

Secretory factors of human chorion-derived stem cells enhance activation of human fibroblasts

Background aimsWound healing remains a principal challenge in modern medical science. Chorion-dervied stem cells (CDSCs), isolated from human placenta, have largely been overlooked, and few studies on their potential in wound healing have been conducted. In this study, we investigated the functional characteristics of CDSCs compared with adipose-derived stem cells (ASCs) on human fibroblasts (HFs). MethodsWe analyzed CDSCs by means of flow cytometry to confirm their mesenchymal stromal cell characteristics. We then evaluated the paracrine effects of CDSCs on HFs in a co-culture system and focused on fibroblast proliferation, migration and collagen synthesis. To explore the potential of CDSCs in wound healing, CDSC- and ASC-secreted factors were compared by use of a cytokine antibody array. ResultsCDSCs had morphology similar to MSCs and expressed a mesenchymal stromal cell phenotype. HF proliferation and migration increased more than 5-fold when co-cultured with CDSCs. Furthermore, Western blot and reverse transcription–polymerase chain reaction analysis showed that expression of collagen (types I and III) in fibroblasts was upregulated 2-fold when co-cultured with CDSCs. Cytokine array results of CDSC-conditioned medium and ASC–conditioned medium revealed the presence of growth factors known to influence wound healing, including interleukin -6, interleukin -8, monocyte chemotactic protein 1 and regulated on activation, normal T cells expressed and secreted. ConclusionsOur data demonstrated that CDSCs are functionally similar to ASCs, promote HF activation, and secrete growth factors that influence wound healing. Therefore, we suggest that CDSCs are potentially applicable in wound healing.

Pro‐angiogenic potential of human chorion‐derived stem cells: in vitro and in vivo evaluation

Human chorion-derived stem cells (hCDSC) were previously shown to demonstrate multipotent properties with promising angiogenic characteristics in monolayer-cell culture system. In our study, we investigated the angiogenic capability of hCDSC in 3-dimensional (3D) in vitro and in vivo angiogenic models for the purpose of future application in the treatment of ischaemic diseases. Human CDSC were evaluated for angiogenic and endogenic genes expressions by quantitative PCR. Growth factors secretions were quantified using ELISA. In vitro and in vivo vascular formations were evaluated by histological analysis and confocal microscopic imaging. PECAM-1+ and vWF+ vascular-like structures were observed in both in vitro and in vivo angiogenesis models. High secretions of VEGF and bFGF by hCDSC with increased expressions of angiogenic and endogenic genes suggested the possible angiogenic promoting mechanisms by hCDSC. The cooperation of hCDSC with HUVECS to generate vessel-like structures in our systems is an indication that there will be positive interactions of hCDSC with existing endothelial cells when injected into ischaemic tissues. Hence, hCDSC is suggested as the novel approach in the future treatment of ischaemic diseases.

Human chorion-derived stem cells: changes in stem cell properties during serial passage

Fetal membrane from human placenta tissue has been described as a potential source of stem cells. Despite abundant literature on amnion stem cells, there are limited studies on the stem cell properties of chorion-derived stem cells. The main aim was to determine the stemness properties of serial-passaged human chorion-derived stem cells (hCDSC). Quantitative polymerase chain reaction (PCR) was performed to reveal the following stemness gene expression in serial-passaged hCDSC: Oct-4, Sox-2, FGF-4, Rex-1, TERT, Nanog (3), Nestin, FZD-9, ABCG-2 and BST-1. Cell growth rate was evaluated from passage (P) 1 until P5. The colony-forming unit-fibroblast (CFU-F) frequency of P3 and P5 cells and multilineage differentiation potential of P5 cells were determined. The immunophenotype of hCDSC was compared using the surface markers CD9, CD31, CD34, CD44, CD45, CD73, CD90, CD117, HLA-ABC and HLA-DR, -DP and -DQ. Immunostaining for trophoblast markers was done on P0, P1, P3 and P5 cells to detect the contamination of trophoblasts in culture, while chromosomal abnormality was screened by cytogenetic analysis of P5 cells. The surface markers for mesenchymal lineage in hCDSC were more highly expressed at P5 compared with P3 and P0, indicating the increased purity of these stem cells after serial passage. Indeed, all the stemness genes except TERT were expressed at P1, P3 and P5 hCDSC. Furthermore, human chorion contained high clonogenic precursors with a 1:30 CFU-F frequency. Successful adipogenic, chondrogenic and osteogenic differentiation demonstrated the multilineage potential of hCDSC. The karyotyping analysis showed hCDSC maintained chromosomal stability after serial passage. hCDSC retain multipotent potential even at later passages, hence are a promising source for cell therapy in the future.