Relevance of syndecan-1 in the trophoblastic BeWo cell syncytialization

Citation Prakash GJ, Suman P, Gupta SK. Relevance of syndecan‐1 in the trophoblastic BeWo cell syncytialization. Am J Reprod Immunol 2011; 66: 385–393 Problem To investigate the role of syndecan‐1 in the differentiation of the BeWo cells into syncytiotrophoblast. Method of study BeWo cells were stimulated with forskolin to form syncytia, and the expression of syndecan‐1, desmoplakin I+II, human chorionic gonadotrophin (hCG) and angiogenesis‐associated factors was analyzed. Syndecan‐1 was silenced by siRNA to evaluate its involvement in the forskolin‐mediated syncytia formation. Results Treatment of the BeWo cells with forskolin led to a significant increase in the syncytia formation. It was associated with an increase in the expression of syndecan‐1 with a concomitant decrease in the expression of desmoplakin I+II. Forskolin treatment of the BeWo cells also led to an increase in the secretion of soluble endoglin, whereas no change was observed in the soluble fms‐like tyrosine kinase‐1. Silencing of the syndecan‐1 expression in BeWo cells led to a significant decrease in cell fusion both in the presence and in the absence of forskolin. It was associated with a significant decrease in hCG level in the conditioned medium. Conclusion Syndecan‐1 is up‐regulated in BeWo cells during differentiation and its silencing inhibits syncytialization and thus could be a useful biomarker for syncytiotrophoblast formation.     

Chromosomal variation in lymphoblastoid cell lines

Tens of thousands of lymphoblastoid cell lines (LCLs) have been established by the research community, providing nearly unlimited source material from samples of interest. LCLs are used to address questions in population genomics, mechanisms of disease, and pharmacogenomics. Thus, it is of fundamental importance to define the extent of chromosomal variation in LCLs. We measured variation in genotype and copy number in multiple LCLs derived from peripheral blood mononuclear cells (PBMCs) of single individuals as well as two comparison groups: (1) three types of differentiated cell lines (DCLs) and (2) triplicate HapMap samples. We then validated and extended our findings using data from a large study consisting of samples from blood or LCLs. We observed high concordances between genotypes and copy number estimates within all sample groups. While the genotypes of LCLs tended to faithfully reflect the genotypes of PBMCs, 13.7% (4 of 29) of immortalized cell lines harbored mosaic regions greater than 20 megabases, which were not present in PBMCs, DCLs, or HapMap replicate samples. We created a list of putative LCL-specific changes (affecting regions such as immunoglobulin loci) that is available as a community resource.     

Chromosomal breakpoints in cholangiocarcinoma cell lines

Little is known about the genetics and biology of cholangiocarcinoma (intrahepatic bile duct carcinoma). Only three human bile duct carcinoma cell lines have been described in the literature. We present the first detailed cytogenetic analysis of two cell lines; a new cell line designated PCI:SG231, established in our laboratory, and RPMI-7451, a previously established cell line. Both lines had highly aneuploid karyotypes with complex rearrangements including marker chromosomes. PCI:SG231, harvested after 50 days in culture, had a modal and median chromosome number of 65, and many cells contained double-minute chromosomes. RPMI-7451 had a modal and median chromosome number of 67. C-banding confirmed the presence of dicentric chromosomes in PCI:SG231. Q-banding confirmed the absence of the Y chromosome in PCI:SG231 and the presence of a der(1)t(Y;1)(q11;p11) chromosome in RPMI-7451. Numerical abnormalities common to both lines included trisomies 2, 5, 11, and 20. Chromosomes 1, 5, 7, and 12 were most commonly involved in structural abnormalities in both lines. Consistent chromosomal breakpoints included 7q22 and 12p11-12. PCI:SG231 was tumorigenic in immunosuppressed nude mice and was histologically similar to the original tumor. Additional cholangiocarcinoma cell lines are being developed to continue the study of the genetics and cell biology of this disorder.     

Chromosomal rearrangement in choriocarcinoma cell lines

Malignant trophoblastic cells from four choriocarcinoma cell lines were evaluated in detail using Q, G, and C banding at various passages. The modal chromosome numbers for BeWo, DoSmi, ElFa, and Jar were 73, 71, 77, and 72, respectively. All the four tumor cell lines exhibited extensive chromosomal rearrangements with several consistent marker chromosomes in each. The majority of these markers have not been previously recognized in this malignancy. Rearrangements of chromosomes 1, 7, 9, 10, and 12 were noted in all four cell lines, but abnormalities of chromosomes 1 and 12 were not consistently present in ElFa and Jar, respectively. Telomeric associations were observed in two cell lines involving chromosomes 11 and 21 as well as chromosomes 3 and 12, resulting in two consistent marker chromosomes. A total of 86 breakpoints were involved in the consistent rearrangements observed in all four cell lines. Most of these breakpoints were located on chromosomes 1, 3, 9, 13, 12, 7, and 21, in order of frequency.     

Pluripotent stem cell lines

The derivation of human embryonic stem cells 10 years ago ignited an explosion of public interest in stem cells, yet this achievement depended on prior decades of research on mouse embryonic carcinoma cells and embryonic stem cells. In turn, the recent derivation of mouse and human induced pluripotent stem cells depended on the prior studies on mouse and human embryonic stem cells. Both human embryonic stem cells and induced pluripotent stem cells can self-renew indefinitely in vitro while maintaining the ability to differentiate into advanced derivatives of all three germ layers, features very useful for understanding the differentiation and function of human tissues, for drug screen and toxicity testing, and for cellular transplantation therapies. Here we review the family of pluripotent cell lines derived from early embryos and from germ cells, and compare them with the more recently described induced pluripotent stem cells.     

B cell hyperactivity in autoimmune continuous B cell lines

Generalized increase in immunoglobulin secretion, which is a prominent feature of autoimmune diseases, may be due to abnormal T cell regulation, intrinsic abnormality of B cells, or both. To investigate this question we developed nonmalignant continuous B lymphocyte lines from 20-week-old BWF1 mice and compared their growth and immune response to that of BALB/c mice cell lines. The B cell lines contain less than 1% T cells and macrophages and require growth factors from phytohemagglutinin-stimulated EL-4 lymphoma (GF) or recombinant interleukin 4 for continuous growth. No antigens or mitogens are required for growth. In the presence of 20% GF (which is optimal for BALB/c cell growth and immune function) spontaneous growth of BWF1 B cells, and spontaneous entry into G1, was similar to that of BALB/c B cells. With concentrations of GF and anti-mu which were optimal for BALB/c, the growth and immune response of isolated BWF1 B cells are no different from those of BALB/c controls, but at suboptimal doses of GF there is a significant increase of both spontaneous immunoglobulin secretion and response to anti-mu in BWF1 B cells. Thus, these autoimmune B cells are more sensitive to the effects of both T cell factors and immunoglobulin receptors stimulation.     

Microsatellite instability in lymphoid leukemia and lymphoma cell lines but not in myeloid leukemia cell lines.

Microsatellite instability (MSI) represents a defect in the DNA mismatch repair system and has been shown to take part in the genesis and/or progression of several human malignancies. In hematological malignancies, the relevance of MSI has been a matter of controversy. Therefore, 29 microsatellite loci were examined for MSI in 57 leukemia and lymphoma cell lines by PCR analysis. Ladder formation of bands representing MSI was observed at multiple loci in 6 of 24 lymphoid leukemia/lymphoma cell lines and in 0 of 33 myeloid leukemia cell lines. Analysis for the BAT-26 and BAT-25 loci confirmed the presence of MSI in five of six lymphoid cell lines exhibiting ladder formation of bands. Thus, at least 5 out of 24 (21%) lymphoid leukemia/lymphoma cell lines were considered as being MSI-positive. These results indicate that MSI contributes to the development of lymphoid but not to myeloid malignancies.     

Gestational trophoblastic disease, Villous gestational trophoblastic disease

Gestational trophoblastic disease (GTD) represents a wide range of clinical and pathological distinct entities. The villous forms of GTD includes developmental disorders of the placental tree, like blighted ovum, embryonal, partial and complete moles. The risk of persistent GTD is estimated of 2-14% in partial and up to 50% in complete moles. So, the morphologic differentiation between the different entities of villous forms of GTD is clinical very important. Sometimes, early forms of complete moles (up to 12th weeks of gestation) may represent diagnostic problems, even in the diagnosis of regressive alterations of the placental villous tree after intrauterine retention.     

Serological analysis of cell surface antigens in choriocarcinoma cell lines

Major histocompatibility complex (MHC) antigens as well as blood group related antigens were investigated in four newly established gestational choriocarcinoma cell lines (NaUCC-1, -2, -3, and -4) using protein A and immune adherence assays. Antibodies to both monomorphic determinants of HLA class I antigens and beta 2 microglobulin reacted with all of the choriocarcinoma cell lines at levels equal to or much greater than SCH, which was teratoid choriocarcinoma cell line. Antibodies to polymorphic determinants not only of the patient's HLA type, but also of the husband's type, reacted to NaUCC-1 and -2. These results indicated that all four newly established choriocarcinoma cell lines express class I HLA antigens. However, we could not demonstrate expression of class II antigens. No expression of blood group A and B antigens could be established, and binding of antibody to Rho(D) antigens was only positive in the NaUCC-1 cell line. These results suggest that some choriocarcinoma cell lines actually express alloantigens and that choriocarcinomas have the character of transplanted tumors that have activities to induce transplantation immunity of the patients.     

Interferon production in murine macrophage-like cell lines

Five murine macrophage (M phi)-like cell lines were examined to determine their suitability for the characterization of M phi interferons (IFNs). The J774A.1, RAW 309 Cr.1, and RAW 264.7 cell lines produced 30-800 international units (IU)/10(6) cells when treated with 5-200 micrograms of bacterial lipopolysaccharide (LPS). No IFN was detected in LPS-treated P388D1 or PU5-1.8 cell cultures. All cell lines produced IFN when inoculated with Newcastle disease virus (NDV); however, only 15 IU/10(6) cells of acid stable IFN were produced in PU5-1.8 cell cultures in comparison to 4.2 X 10(3)-1.7 X 10(4) IU/10(6) cells in the other cell lines. Most of the IFN was produced within 4 h in LPS-treated cell cultures and within 12 h in NDV-infected cell cultures. All IFNs were stable at pH 2.0 and were neutralized with antiserum against mouse L cell IFN. These cell lines appear competent for use in studying the synthesis, molecular weights, and regulatory functions of M phi IFNs.     

Development of tumor cell lines

A number of techniques are now in use in attempting to establish cell lines from human solid tumors. Although short-term cultures are readily attained, relatively few become established as permanent cell lines, especially from primary carcinomas where metastasis has not already occurred and from such organs as the breast, prostate, and pancreas. With recent advances in techniques and in our knowledge of growth factors, however, these difficulties may be resolved in the near future.     

Identification of insect cell lines

Identification of 10 cell lines of 6 insect species received by the cell culture collection of Molecular Biology Institute from different research institutions of the USSR was carried out by karyological analysis and determinations of the spectrum of isoforms of the following enzymes: glucose-6-phosphate dehydrogenase (G-6-PDH), lactate dehydrogenase (LDH), isocitrate dehydrogenase (ICDH), and superoxide dismutase (SOD). Nine out of the 10 cell lines examined were shown to be identical to those of gypsy moth. Possible ways of cell contamination with other cells in long-term cultivation and preparation of new lines were determined.     

HLA expression in hepatocellular carcinoma cell lines

The present study undertook to investigate the biological significance of human leucocyte antigen expression in hepatocellular carcinoma and to elucidate the role of potential modulating agents on human leucocyte antigen expression. These studies used several hepatic tumour-derived cell lines as in vitro model systems. The cell lines included PLC/PRF/5 (Alexander cell line), Hep3B. HepG2, TONG PHC, HA22T/VGH, HA59T/VGH and Mahlavu, The cell lines K562 and Raji were used as negative and positive controls, respectively. K562, a B tymphoid-derived cell line, was shown to express negligible amounts of human leucocyte anligens, while Raji, an erythromyeloid-derived cell line, expressed both class I and class II human leucocyte anligens as well as their respective invariant chains, β₂-niicroglobulin and Ii. Using an ELISA, experiments performed on these cell lines confirmed the natural expression of class I and class II antigens by the HA22T/VGH and HA59T/VGH cell lines, whereas PLC/PRF/5 displayed class II surface antigens only. The effects of modulating agents such as interferon-gamma sodium bulyrate and clofazimine on human leucocyte antigen expression were investigated using the HA22T/VGH, HA59T/VGH and TONG PHC cell lines. These agents increased class 1 and class II human leucocyte antigen expression on HA22T/VGH and TONG PHC cells, but had no effect on the HA59T/VGH cell line. The results suggest a potential use for these agents as modulators of human leucocyte antigen expression by human heptocellular cell lines.     

Four new human germ cell tumor cell lines

Four new human tumor cell lines, NCC-EC-1, NCC-EC-2, NCC-EC-3, and NCC-IT, were derived from different germ cell tumors established in vitro. On the basis of morphologic studies of cell cultures and nude mice xenografts, it was concluded that NCC-EC-1, and NCC-EC-2 cell lines are equivalent to developmentally nullipotent embryonal carcinoma; cell line NCC-EC-3 showed trophoblastic differentiation, whereas the NCC-IT cell line was composed of developmentally pluripotent cells capable of somatic and extraembryonic differentiation. Nude mouse-xenografts of NCC-IT contained foci of embryonal carcinoma, yolk sac tumor, immature somatic tissues, and trophoblastic giant cells indicating that this cell line is indeed developmentally pluripotent. We conclude that human embryonal carcinoma cell lines may be developmentally nullipotent, show restricted capacity for differentiation, or be developmentally pluripotent.