In vitro cultivation

Santos RPD, MariathDeAraujoJE. A simple method for fixing dehydrating pollen tubes cultivated in vitro for transmission electron microscopy. Histochemistry 1997 76 315-319. 

J.G. Torrey. Isolation and cultivation in vitro of the actinomycete causing root nodulation in Comptonia. Science 199,899-902 (1978). 

Particles from cationic lipids may also be useful for antisense therapy of skin disease — a nontoxic increase in the oligonucleotide uptake by cultivated keratinocytes and a sebocyte cell line has been reported [66]. Moreover, cationic dendri-mers also efficiently transfer reporter gene DNA to human keratinocytes cultivated in vitro. In the skin of hairless mice, in vivo transfection was possible with complexes, yet reporter gene expression was localized to perifollicular areas. Transfection, however, failed with the naked plasmid. For prolonged contact, biodegradable membranes coated with dendrimer/DNA complexes were used [67]. This hints at a follicular uptake of these complexes and indicates that gene transfection also may be possible with human skin, which has a thicker stratum comeum compared with mouse skin (eight to ten vs. two to three layers [58]). 

Implants made of TCP and, to some extent, also of HA are partly resorbed after the implanta-tion. This may take place cellularly, in which case the material will be incorporated into the normal physiological process of making and breaking down bone. This phenomenon promotes the formation of new bone material and the strong connection between the rest of the implant and the living tissue. By immersing the bio-material into the patient s osteoblasts (= self bone cells) which have been cultivated in vitro, we can stimulate these processes. 

The first tissue engineering product to be approved targeted the treatment of burns and consisted of keratinocytes cultivated in vitro, which form a tissue that is later transplanted to the patient. Beyond this product, others are already being commercialized for a wide range of applications. Examples are Carticel (marketed by Genzyme), which consists of chondrocytes employed in the treatment of cartilaginous defects caused by acute or repetitive traumas Apligraf (Novartis), used in the treatment of venous ulcer and DACS SC (Dendreon), utilized for reconstitution of the immune system after chemotherapy. 

Various animal cell lines can be cultivated in vitro, such as cardiac cells, fibroblasts, smooth muscle cells, endocrine cells (such as pituitary, adrenal, and pancreatic cells), epithelial cells (such as liver, mammary, lung, and kidney cells), tumor cells (such as melanocytes), nervous system cells (such... 

Cells can be cultivated in vitro after a tissue is dissected and totally or partially disaggregated by enzymic treatment. Cell, explants, or organ cultures can be established depending on the degree of disaggregation of the tissue, as shown in Figure 2.2. 

Most cell lines present a wide tolerance range to osmotic pressure. Given the osmolality of human plasma (290 mOsm/kg), it may be reasonable to assume that this is an optimal value for human cells cultivated in vitro, although it can be different for cells from other species. As a general rule, the optimal osmolality range for mammalian cells in culture is from 260 to 320 mOsm/kg, while for insect cells higher values are optimal, from 340 to 390 mOsm/kg. 

Courduroux, J.-C., Gendraud, M., and Teppaz-Misson, M., Action comparee de l acide gibberellique exogene dt du froid sur la repousse et la levee de dormance de tubercules de Topinambour (Helianthus tuberosus L.) cultives in vitro, Physiol. Veg., 10, 503-514, 1972. 

Durst, F. and Duranton, H., Quelques aspects de l influence de la limiere sur le metabolisme de l arginine dans les tissus du Topinambour cultives in vitro, Physiol. Veg., 4, 283-298, 1966. 

Gautheret, R.J., Action de la lumiere et de la temperature sur la neoformation de racines par des tissus de Topinambour cultives in vitro, C. R. Acad. Sci. Paris, 252, 2791-2796, 1961a. 

Gautheret, R.J., Sur l interaction des principaux facteurs de laneoformation des racines par les tissus de Topinambour cultives in vitro, in 90eme Congres des Society Savantes, Nice, France, 1965, pp. 355-368. 

Gautheret, R.J., Action de variations de temperature sur la rhizogenese des tissues de Topinambur cultives in vitro, in Les Cultures de Tissus de Plantes, No. 193, Colloques Intemationaux du CNRS, Paris, 1971, pp. 187-199. 

Gendraud, M. and Pre vot, J.C., Mesure in situ de Tincorporation d un precurseur dndogene dans les ARN des tubercules de Topinambour cultives in vitro en fonction de leur etat de dormance, Etude critique et exemple de resultats, Physiol. Veg., 11, 417 -27, 1973. 

Durst, F. and Duranton, H., Phytochrome and phenylalanine ammonia-lyase in Helianthus tuberosus tissues cultivated in vitro, C. R. Seances Acad. Sci. D, 270, 2940-2942, 1970. 

Constabel, F. and Haala, G. (1968) Recherches sur la formation de pigments dans les tissus de betterave fourragere cultive in vitro. Colloques Nationaux Centre National de la Recherche Scientifique, Paris, pp. 223-9. 

Ruano GU D. The influence of thalidomide on the development of chick embryos cultivated in vitro. Acta Anat (Basel) 1967 66(2) 226-37. 

Although in principle the cells producing these might be isolated and cultivated in vitro, the quantities produced still be small and the costs of cultivaticm are hi. ... 

Multiple myeloma is a disease resulting from the malignant transformation of a unique clone of plasmocytes which proliferate in the bone marrow. These cells synthesize and secrete a monoclonal immunoglobulin. They may be cultivated in vitro indefinitely. Fusion with antigen-activated B lymphocytes gives a... 

The artificial virulentation occurred as follows. Strain F. 97 was isolated from sclerotia grown on plants of Paspalum distichum, collected at Tivoli (Rome) and identified and classified as Claviceps paspali Stevens and Hall. Embryos of Rosen 4 n rye were inoculated, before germination, with the strain F. 97 and then cultivated in vitro. The new virulented subspecies were isolated from sclerotia obtained on said embryos. 

As regards the production of alkaloid derivatives of lysergic acid, the present invention is not limited to the use of the described strains, but comprises also the mutants thereof, which may be obtained, e.g., by means of either a selection or a mutation by the action of U.V. rays or Roentgen rays or any other mutagenous substance or, particularly, by artificial infection of either embryos or grasses cultivated in vitro or plants of grasses cultivated both in vivo or in vitro and the said mutants are to be included in the definition of a new strain of Claviceps paspali Stevens and Hall. 

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