Extended organ culture of tumor cell lines with chick embryonic tissues

Summary An organ culture method has been developed in which human tumor cells and chick embryonic tissues are cocultured on agar platforms surrounded by replaceable liquid medium. This simple system permits such cultures to remain viable for up to 60 d; the extended culture period allows the tumor cells to invade, organize, and differentiate and to form microscopic tumors in embryonic tissues similar to tumors formed by the tumor cells in vivo in nude mice. These microtumors can also be examined for tumor-associated antigen expression by conventional immunoperoxidase techniques.     

Nonadherent,stationary organ culture of human respiratory mucosa

Summary Fragments of human adenoid tissue were grown in a tissue culture system where all artificial tissue handling, except for the explanation procedures and tissue culture condition in itself, was excluded. The fragments formed spheroids that were covered by a pseudostratified, ciliated epithelium. The epithelium rested on a basement membrane. The central parts of the spheroids consisted of fibroblasts and collagen fibers. This structure remained unchanged for the 40 days the fragments were observed in culture.     

Isolation and culture of heart cells from the European flat oyster,Ostrea edulis

The present study reports a culture technique for heart tissues of the European flat oyster,Ostrea edulis. Heart tissues of flat oysters were dissociated by a trypsin-EDTA treatment and a mechanical action in a Dounce-type homogeneizer. Then, dissociated cells were cultured in three different synthetic media, in pure sea water or in sea water mixed with sterile filtered Japanese oyster,Crassostrea gigas, hemolymph. All these media were supplemented with 10% of fetal bovine serum. Cultures were grown at 20°C in previously Poly-D-Lysin coated flasks or culture wells. The optimized medium, 3% L15 medium (w/v) in sterile sea water mixed with Japanese oyster hemolymph (1:1) and supplemented with 10% of fetal bovine serum gave the best result. Morphological characterization for the cardiac cultured cells was performed. Thus, cell monolayers of dissociated heart tissues consisted essentially of hemocytes and large granular pigmented cells.     

Brain tissue in nonadherent stationary organ culture

Summary In vitro growth of histiotypic brain tissue fragments is described. The tissue is not exposed to artificial mechanical forces or chemical agents. It does not adhere to or interact with any underlying surface or surrounding material.     

A method for in vitro cell culture of superficial bovine uterine endometrial epithelium

Summary Bovine epithelial cells were obtained for culture from the uterine endometrium of adult, cyclic cattle. Using the procedures described herein, cell-specific monolayers of uterine epithelial cells developed rapidly in culture and maintained a good level of viability for seven to eight subcultures. In addition, frozen-thawed uterine epithelial cells also maintained a respectable level of viability during postthaw subculture. Patterns of cell growth for uterine epithelial cells were determined by a growth curve. A growth curve of fresh epithelial cells over an 8-day interval revealed a short lag phase (24 h) followed by a log growth phase for 5 days and then a stationary phase starting on Days 6 or 7 of incubation. This method for isolation and culture of uterine epithelial cells provides a potential model for evaluating uterine epithelial cell secretory capacity during the estrous cycle. This culture system may offer benefits for in vitro culture of bovine embryos.     

猪眼小梁组织眼前节灌流培养和组织块培养

目的了解猪眼小梁组织的灌流眼前节培养和组织块培养在青光眼研究中的应用。方法采用恒速眼前节灌流培养和组织块培养法培养猪眼小梁组织,用光学显微镜观察不同培养方法保存的小梁组织的状态。结果恒速(0.1mL/h)灌流猪眼前节,眼压可以稳定在正常眼压范围(10~12mmHg);在一定范围内,随着流速增加眼压能逐步升高。灌流培养的组织大体形态保留完好,组织层次清晰,细胞状态好。组织块原位培养的小梁组织,周边组织结构保留完好,细胞状态尚可,但是小梁网间隙不如灌流培养的小梁网间隙饱满。结论灌流眼前节培养法可作为短期高眼压模型,更接近小梁网的生理状态,小梁网的正常充盈是维持其功能和结构的必要条件。     

Bovine tongue organ culture assay for tumorigenicity

Summary Bovine tongue purchased from public retail food distribution centers was used as the substrate onto which tumor cells were inoculated in organ culture. The tongue could be preseved at 4°C for several weeks before use in the assay, and it served as an excellent substrate on which tumor cells grew as well as into which they invaded. The system mimics the intramuscular inoculation of tumor cells in vivo, but without the complications of immunologic rejection mechanisms.     

cDNA cloning and molecular identification of the major oyster allergen from the Pacific oyster Crassostrea gigas

BACKGROUND: Shellfish is one of the most common food allergens. Despite the recent cloning and molecular identification of the major heat stable crustacean allergens in shrimp, lobster and crab, there have been no similar studies on molluscs to which a significant portion of populations allergic to shellfish are also hypersensitive. Recent biochemical evidence suggests that tropomyosin is also an allergen in molluscs, but data on the molecular cloning, nucleotide sequencing, expression and IgE binding to mollusc tropomyosin are lacking. OBJECTIVE: This study was undertaken to clone, identify and determine the primary structure of a major IgE-reactive mollusc allergen in oyster at the DNA and protein level. METHODS: We constructed an expression cDNA library from the Pacific oyster Crassostrea gigas. This library was screened for IgE binding clones using sera from 15 subjects with a well-documented history of type I hypersensitivity reactions to oysters. An IgE reactive clone was selected and sub-cloned into plasmids for nucleotide sequence determination and expression in E. coli. RESULTS: We identified a 1.3-kb cDNA designated as Cra g 1.03. Expression of Cra g 1.03 in plasmid vector pGEX produced a 59-kDa recombinant fusion protein reactive to the IgE antibodies from patients with oyster allergies but not non-allergic controls. Cra g 1.03 has an open reading frame of 233 amino acids and demonstrates marked similarity in amino acid composition and peptide sequence with mollusc and crustacean tropomyosins. Absorption of oyster allergic sera with Cra g 1.03 totally removed IgE reactivity to oyster extract. Moreover, absorption of allergic sera with recombinant shrimp tropomyosin (Met e 1), lobster tropomyosin (Pan s 1) and crab tropomyosin (Cha f 1) removed most of the IgE reactivity to Cra g 1.03. CONCLUSION: Cra g 1.03 is the first oyster allergen identified at the molecular level. Nucleotide and amino acid comparison shows that this protein is the oyster tropomyosin.     

A method for the organ explant culture of fish kidney tubules

Summary A method for long-term organ explant culture of isolated fish kidney tubules from two teleost species is provided. Tubules maintained normal structures for 8 weeks when cultured in supplemented CMRL-1066 medium in controlled atmosphere chambers.     

High oxygen atmosphere improves human follicle development in organ cultures of ovarian cortical tissues in vitro

BACKGROUND: Obtaining mature human follicles from cultured ovarian tissue may be beneficial for clinical use for women who wish to preserve fertile competence. However, the methodology of culture such as culture condition and gas atmosphere has not been well established in humans. Therefore, we investigated the effect of oxygen concentration in organ culture in order to establish an ovarian tissue culture method. METHODS: Ovarian tissue was obtained from 26-35-year-old women undergoing removal of a benign tumor (n = 12) or caesarean section (n = 16). The ovarian cortical tissues were cultured on a cell culture insert for 15 days under high (100%) and low (air, 20%) oxygen concentrations and then inspected for follicle development with light and electron microscopy. Estradiol and progesterone concentrations in the medium during culture were measured. RESULTS: The ultrastructure and the function of hormone secretion in the cultured tissues were well preserved after organ culture. The follicles developing under high oxygen were larger and more matured than those developing under low oxygen (P < 0.05). CONCLUSIONS: Human ovarian tissues can be cultured for 15 days under high oxygen concentration with the organ culture system used here. This technique could make it possible to utilize ovarian tissue for preservation of reproductive competence in cancer patients.     

用于组织工程化培养生物反应器的研究进展

生物反应器是组织工程研究与临床应用的重要工具之一 ,近年来一直受到国内外学者和企业的广泛关注。本文系统地介绍了各种用于组织工程化培养生物反应器的研究现状。由于生物反应器的机械性能、传质以及流体剪应力等因素对培养组织的形态和功能有很大的影响 ,因比 ,深入研究和开发新型生物反应器对组织工程的研究和今后临床的应用都有着十分重要的意义     

Biological activity of hepatitis B antigens in cell culture

Direct interference by purified hepatitis B surface antigen or virus particles was not demonstrated in tissue culture. Significant levels of interferon were not induced. The surface antigen did not block the adsorption of other viruses.     

Hematopoietic stem cell production in vitro: Washings from embryonic liver organ cultures

Hematopoietic cells, including splenic colony-forming units (CFUs) can be periodically washed off an embryonic liver organ culture in the course of 4 weeks. Under the conditions of culture used this operation does not substantially reduce the numer of CFUs in the culture. The washings can thus be used to increase the yield of CFUs from the culture.Laboratory of Bone Marrow Culture and Transplantation, Central Research Institute of Hematology and Blood Transfusion, Ministry of Health of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR N. A. Fedorov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 87, No. 3, pp. 281–282, March, 1979.     

Intact organ culture of murine metanephros

Summary A murine metanephric whole-organ culture system is described which maintains normal structural relationships of epithelial and mesenchymal elements and undergoes advanced organotypic differentiation in vitro. Manipulation of the culture conditions permits study of the developmental, immunologic, and biochemical aspects of normal and abnormal renal organogenesis.     

Tissue culture of bovine subcommissural organ

Tissue of the secretory, glial subcommissural organ (SCO) of adult, male cattle was cultured in serum-free medium for 70 days in vitro. Only minor alterations in the histoarchitecture and the cytology of the explanted SCOs could be observed by light and electron microscopy. Light- and electron-microscopic immunocytochemical investigations with an antiserum raised against bovine SCO secretory proteins revealed intra- and extra-cellularly localized immunoreactive material in tissue sections of SCO explants cultured up to 69 days in vitro. An indirect competition enzyme-linked immunosorbent assay (ELISA) was developed to detect minor quantities of SCO secretory products. By means of this assay, approximately 35 ng RF protein per ml was detected in culture medium supernatants conditioned for 3 days in SCO tissue cultures at 3, 38, and 69 days in vitro. These studies demonstrate that the bovine SCO can maintain its secretory activity throughout long periods in vitro.