Characteristics of amniotic fluid derived stem cells with trisomy 21 / 대한산부인과학회지

OBJECTIVE: To assess molecular markers of amniotic fluid derived stem cells (AFSCs) in aspects of increased neurological deficit in Down syndrome. METHODS: Amniotic fluid samples through amniocentesis for prenatal diagnosis from four mid trimester pregnancies; by routine chromosomal analysis, two of them were trisomy 21 (Down syndrome) and others were normal, were selected after informed consent. Cells from two-stage culture protocol were assayed; morphology through phase contrast microscopy, chromosomal analysis, reverse transcriptase-polymerase chain reaction and Western blot analysis. RESULTS: AFSCs were highly proliferative in subcultures and most of them were mononuclear, fibroblast-like, fusiform cells. There were also a few ovoid cells. The chromosomal analysis of amniotic fluid stem cells was identical to that of amniotic fluid cells. Two of four samples were 47,XX,+21, others were 46,XX. Of the proteins related to Down syndrome, the expression of S100beta were increased in AFSCs of Down syndrome, COL6A1 (Collagen IV, alpha 1) was down-regulated in them and insulin like growth factor binding protein-1 was expressed in all AFSCs. Stem cell markers were expressed heterogeneously. Oct4 (POU5F1), nanog, and SOX2 (sex determining region Y) were expressed in both groups. But c-Kit was not expressed in AFSCs of Down syndrome. The neural cell marker, neuron specific enolase was detected in both groups. Other neural cell markers, microtubule associated protein 2, glial fibrillary acidic protein were undetectable in ASFCs of Down syndrome. Bcl-2 gene family proteins related with apoptosis were assayed. The expression of Bcl-XL was increased in Down syndrome more than in normal pregnancy. Bcl-2 and BID were expressed in all AFSCs and Bax was down-regulated in Down syndrome. CONCLUSION: AFSCs are an excellent choice for many future tissue engineering strategies and cell based therapies. Analysis of molecular features of AFSCs from normal and Down syndrome will provide the basis of further experimental study.

Characterization and differentiation into adipocytes of mesenchymal stem cells (MSCs) from human adipose tissue and amniotic fluid / 대한산부인과학회지

OBJECTIVE: Mesenchymal stem cells (MSCs) are potentially very useful for regenerative and reparative medicine as well as therapeutic possibilities. The aim of this study is to examine the ability of ADSCs and AFSCs to be phenotypically and functionally differentiated into adipocyte and to determine the appropriate stem cell source and conditions for efficient adipocyte regeneration. METHODS: Adipogenic differentiation was induced by culturing confluent ADSCs and AFSCs in adipogenic medium for 2~4 weeks. During the differentiation inducing period, we evaluated the successful adipogenesis by performing immunocytochemistry and RT-PCR to detect the lipid producibility and several adipogenic gene expressions including lipoprotein lipase (LPL), peroxisome proliferator-activated receptor gamma2 (PPAR gamma2) and adiponectin. RESULTS: ADSCs and AFSCs are expanded easily in vitro and exhibited a fibroblast-like morphology as previously known in MSCs from bone marrow and a commercial source. Flow cytometric analysis showed that ADSCs and AFSCs expressed several CD marker antigens similar to those observed on bone marrow-derived MSCs. Adipogenic induction of ADSCs and AFSCs resulted in the extended cell morphology, intracellular staining of an established lipid dye Oil Red O, and expression of adipocyte-specific genes. CONCLUSION: Both ADSCs and AFSCs successfully differentiate in vitro into adipogenic cells in the presence of the lineage-specific induction factors although ADSC showed the greater capability. Therefore, the results suggest that ADSCs and AFSCs may be an excellent choice for many future tissue engineering strategies and cell-based therapies.