Identification of cells from fetal bladder epithelium in human amniotic fluid

Summary Cultured human amniotic fluid cells consist of five different types of cytokeratin-positive epithelial cells, E-1 to E-5, differing by their size, growth morphology, and cytokeratin pattern, according to our earlier investigations. Using anticytokeratin antibodies in indirect immunofluorescence (IIF) microscopy, we show in this study that cultured urine cells contain four of the cell types found in amniotic fluid. In addition, we used two urothelium-specific antibodies, anti-UMA and anti-Las-86, in combination with cytokeratin antibodies to distinguish urothelium-derived cells in amniotic fluid and urine cell cultures. Two of the epithelial cell types were found to express urothelial antigens and thus to originate from the transitional bladder epithelium. These cells were found in 26 of the 33 amniotic fluid cell cultures and in nine of the ten urine cell cultures.     

Structural studies on fetal mucins from human amniotic fluid. Core typing of short-chain O-linked glycans.

Mucins in human amniotic fluid are represented by two distinct molecular species, FM-1 and FM-2, with apparent molecular masses of 700 and 570 kDa, respectively, in SDS/polyacrylamide gradient gels. FM-1 and FM-2 were isolated by preparative SDS/PAGE to apparent homogeneity and subjected to structural studies on their carbohydrate portions. The carbohydrate compositions of the mucin species differed only marginally and exhibited significant amounts of mannose. O-linked core-region glycans on human amniotic mucin-derived pronase-stable glycopeptides were analyzed after reductive beta-elimination and purification on HPLC by a combination of methylation analysis, electron-impact mass spectrometry of permethylated oligosaccharide alditols and fast-atom-bombardment mass spectrometry of acetylated or methylated alditols (positive-ion mode) or alditol-derived neoglycolipids (negative-ion TLC-MS). The primary structures of major monosaccharides to tetrasaccharides have been established which exhibit at their reducing termini core 1, core 2 and core 3 sequences, as follows. [Table; see text]     

Use of genetic markers to certify fetal origin of cultured amniotic fluid cells

Summary Phenotypes of five polymorphic enzymes: red cell acid phosphatase, phosphoglucomutase, esterase D, adenosine deaminase, and 6-phosphogluconate dehydrogenase were determined in extracts of 24 amniotic fluid cell cultures and in the corresponding maternal red cells. Twenty-one of the 24 fetus/mother pairs can be distinguished by at least one of the markers. Thus, polymorphic enzyme markers may be useful in affirmation of fetal origin of cultured cells and to avoid possible diagnostic errors.Work supported by USPHS grants GM 15253 and AI 12617     

Adrenal-like Leydig cells (author's transl)]

Adrenal-like lipoid-rich Leydig cells, which could be found in a cryptorchid testis, were investigated by light and electronmicroscopy. There were nodular and diffuse proliferation of these adrenal-like cells in the interstitium of the testis. Electronmicroscopically these cells are fasciculated and characterized by large liposomes, many tubulovesicular mitochondria, and a large smooth endoplasmatic reticulum. But the presence of crystals of Reinke in these cells underlined their relationship to Leydig cells. The clinical history of this case is characterised by an extreme adipositas (167 kg) and high urinary estrogenexcretion. This excretion could be suppressed with dexamethasone and stimulated with HCG. After orchiectomy estrogen excretion decreased for 4 months and then increased again, after ACTH stimulation performed by reason of adrenal insufficiency. At this time there is no evidence of adrenal tumor; in the contralateral, scrotal testis, spermiogenesis and Leydig cells are without pathologic changes as revealed by biopsy.     

Patterns of enzyme development utilizing cultivated human fetal cells derived from amniotic fluid

Fetal cells obtained from amniotic fluid at various stages of pregnancy were successfully cultivated. Quantitative enzyme analysis and qualitative enzyme analysis, utilizing starch gel electrophoresis, were performed. Increased glucose 6-phosphate dehydrogenase activity associated with a decreased percentage of sex chromatin positive cells were found in cells derived from two 10-week female fetuses. After 6 weeks, these cultures contained normal levels of glucose 6-phosphate dehydrogenase activity and normal numbers of sex chromatin positive cells. Qualitative changes of glucose 6-phosphate dehydrogenase and lactate dehydrogenase were demonstrated.This study is supported by U.S.P.H.S. Grants 1 RO1 HD 02752 and TI AM 5186.     

Stem cells from fetal membranes and amniotic fluid: markers for cell isolation and therapy

Stem cell therapy is in constant need of new cell sources to conceive regenerative medicine approaches for diseases that are still without therapy. Scientists drew the attention toward amniotic membrane and amniotic fluid stem cells, since these sources possess many advantages: first of all as cells can be extracted from discarded foetal material it is inexpensive, secondly abundant stem cells can be obtained and finally, these stem cell sources are free from ethical considerations. Many studies have demonstrated the differentiation potential in vitro and in vivo toward mesenchymal and non-mesenchymal cell types; in addition the immune-modulatory properties make these cells a good candidate for allo- and xenotransplantation. This review offers an overview on markers characterisation and on the latest findings in pre-clinical or clinical setting of the stem cell populations isolated from these sources.