Subject(s)
Chromosomes, Human, Pair 5 , Genetic Markers , Janus Kinase 2/genetics , Myeloproliferative Disorders/genetics , Myeloproliferative Disorders/pathology , Adult , Aged , Aged, 80 and over , Bone Marrow/pathology , Cell Division , Chromosome Deletion , Female , Humans , Male , Middle Aged , Myeloproliferative Disorders/classification , Point MutationABSTRACT
Ever since monoclonal antibodies were produced in 1975 with mouse myeloma cells there has been interest in developing human myeloma cultures for the production of monoclonal antibodies. However, despite multiple attempts, no human myeloma line suitable for hybridoma production has been described. Here we report the derivation of a hypoxanthine-aminopterin-thymidine-sensitive and ouabain-resistant human myeloma cell line (Karpas 707H) that contains unique genetic markers. We show that this line is useful for the generation of stable human hybridomas. It can easily be fused with ouabain-sensitive Epstein-Barr virus-transformed cells as well as with fresh tonsil and blood lymphocytes, giving rise to stable hybrids that continuously secrete very large quantities of human immunoglobulins. The derived hybrids do not lose immunoglobulin secretion over many months of continuous growth. The availability of this cell line should enable the in vitro immortalization of human antibody-producing B cells that are formed in vivo. The monoclonal antibodies produced may have advantages in immunotherapy.
Subject(s)
Antibodies, Monoclonal/biosynthesis , HIV Envelope Protein gp41/immunology , Multiple Myeloma , Tumor Cells, Cultured , Amino Acid Sequence , Cell Line, Transformed , Humans , Hybridomas , Microscopy, Electron , Molecular Sequence DataSubject(s)
Chromosome Aberrations , Chromosomes, Human, Pair 4 , Leukemia, Myeloid, Acute/genetics , Trisomy , Aged , Humans , Karyotyping , MaleABSTRACT
The beta-subunit of human chorionic gonadotrophin (hCG) is coded on chromosome 19 by the beta-hCG-hLH gene cluster. Genomic DNA has been isolated from bladder tumour cell lines which ectopically express beta-hCG. The beta-hCG-hLH gene cluster was probed for possible rearrangement or amplification and cells karyotyped for chromosome 19 abnormalities. No rearrangement or amplification of the gene cluster and no consistent abnormalities of chromosome 19 were found. The expression of beta-hCG by bladder tumours is therefore likely to be the result of altered gene regulation and not a rearrangement or amplification of this gene cluster.
Subject(s)
Chorionic Gonadotropin/genetics , Gene Amplification , Gene Rearrangement , Hormones, Ectopic/metabolism , Luteinizing Hormone/genetics , Peptide Fragments/genetics , Urinary Bladder Neoplasms/metabolism , Cell Line , Chorionic Gonadotropin/metabolism , Chorionic Gonadotropin, beta Subunit, Human , DNA/genetics , Gene Expression Regulation , Humans , Karyotyping , Nucleic Acid Hybridization , Peptide Fragments/metabolism , Tumor Cells, Cultured/metabolismABSTRACT
The maceration technique for the culture of chorionic villi has been applied by St Mary's Hospital and King's College Hospital, to a total of 225 villus aspirates of which 100 were from diagnostic cases. Two hundred and eighteen (96.8 per cent) of these were successfully karyotyped, and chromosome abnormalities were detected in 15 cases. Of the cases with a normal karyotype, 113 were male and 90 were female. Sixty-nine (77 per cent) of the female cultures were demonstrated to be fetal and a further two of these may be verified at delivery. The use of Chang medium was found to accelerate cell growth thereby reducing the interval between sampling and reporting to less than 2 weeks. An unlimited quantity of metaphase spreads was obtained suitable for the application of routine banding techniques, and comparable to those obtained from amniotic fluid culture. Experience with diagnostic cases reinforced our view that culture should always back up direct analysis and this is discussed with particular reference to the occurrence of mosaicism in the trophoblast.
Subject(s)
Chorionic Villi/pathology , Chromosome Aberrations/diagnosis , Chromosome Disorders , Culture Techniques , Female , Humans , Infant, Newborn , Karyotyping , Male , Pregnancy , Pregnancy Trimester, FirstABSTRACT
Aspiration biopsy of trophoblastic villi was performed on 56 patients immediately preceding suction termination of pregnancy, using a malleable metal cannula and ultrasound guidance. Villi were obtained from 47 patients (84%), with sampling success rising to 93% after experience. Immediate complications were noted in 14% of patients and correlated with placental positions situated furthest from the cervical canal. Karyotyping from cultured and/or direct preparations was attempted on the villus samples and was successful in 42. Clarity of the karyotypes obtained from direct preparations in this series was not found to be adequate for diagnostic purposes. A number of practical suggestions which facilitate chorion villus sampling are described.
Subject(s)
Biopsy, Needle/methods , Chorionic Villi/pathology , Prenatal Diagnosis/methods , Chorionic Villi/ultrastructure , Female , Gestational Age , Humans , Karyotyping , Pregnancy , Pregnancy Trimester, First , UltrasonographyABSTRACT
A technique for the culture of chorionic villi for cytogenetic studies is described which is simple, reliable, rapid, and suitable for routine laboratory use. This method eliminates the need for multi-stage and time-consuming procedures inherent in previous techniques which have been developed for this tissue. Cultures suitable for analysis were obtained in an average of sixteen days and the risk of maternal cell contamination was evaluated. Results of cultures initiated from 50 chorion biopsies obtained by transcervical blind aspiration at a weekly termination clinic are presented, and the potential use of this technique as a routine procedure is discussed.
