In vitro differentiation of a cloned bovine mammary epithelial cell

Abstract

Rose, M. T., Aso, H., Yonekura, S., Komatsu, T., Hagino, A., Ozutsumi, K., Obara, Y. (2002). In vitro differentiation of a cloned bovine mammary epithelial cell.? Journal of Dairy Research, 69, (3), 345-355The aim of the study was to establish in vitro a bovine mammary epithelial cell (MEC) clone, able to respond to mitogenic growth factors and to lactogenic hormones. Mammary tissue from a 200-d pregnant Holstein cow was used as a source of MEC, from which a clone was established through a process of limiting dilution. When plated on plastic, the cells assumed a monolayer, cobblestone, epithelial-like morphology, with close contact between cells. Inclusion of IGF-1 and EGF in the media significantly increased the number of cells 5 d after plating. All cells stained strongly for cytokeratin and moderately for vimentin at young and old passage stages, indicating the epithelial nature of this cell clone. When the cells were plated at a high density on a thin layer of a commercial extracellular matrix preparation (Matrigel), lobular, alveoli-like structures developed within approximately 5 d, with a clearly visible lumen. When cells were plated onto Matrigel in differentiation media (containing lactogenic hormones), detectable quantities of ?-casein were present in the media and particularly on the lumen side of the structures. Omission of one of the lactogenic hormones (insulin, prolactin or hydrocortisone) reduced ?-casein release to the limit of detection of the assay used. Lactoferrin was also produced when the cells were plated on Matrigel, again principally on the lumen side of the lobules, though this was independent of the lactogenic hormones. By passage 40, the cells had senesced, and it was not possible to induce ?-casein or lactoferrin production. This study notes the establishment of a functional bovine mammary epithelial cell clone, which is responsive to mitogenic and lactogenic hormones and an extracellular matrix.Peer reviewe

Mammary transmission of caprine arthritis encephalitis virus: a 3D model for in vitro study

“…Then, we observed formation of mammospheres, or acinus-like structures, morphologically similar to those described as deriving from mammary epithelial cell lines [15][16][17] or human breast tissue [18]. This was in accordance with the general property of the cells to develop architectural arrangements when cultured in three-dimensional matrices [19].…”

“…The acinus-like structures were limited by a basal membrane, as expected [15]. The synthesis of alpha-lactalbumin after in vitro stimulation with prolactin established that, as shown in other models [16], a basic physiological function is conserved in vitro. We In our experimental conditions, Matrigel was diluted ½, and its initial structure could be modified by the organization and possible secretion of a cellular matrix between mammospheres (Fig.…”

Establishment of mammary tissue culture: an in vitro model for bovine lactation

“…Accurate replication of in vitro bovine mammary gland function would be of great benefi t to studies on the control of milk synthesis and secretion (Rose et al, 2002). There are a number of possible in vitro mammary incubation methods and some approaches have been developed to quickly and properly evaluate the redox activity and function of the cultured tissue.…”

Modelos para a produção de eritropoietina recombinante humana in vivo e in vitro com vetores plasmideais em ovinos

“…Nas últimas décadas foram estabelecidas diferentes linhagens de células mamárias bovinas (MATITASHVILI; BAUMAN, 1999;LAMMERS et al, 1999;ROSE et al, 2002;WELLNITZ;KERR, 2004e HU et al, 2009. Nessas linhagens foram relatadas expressão protéica de β-caseína (MATITASHVILI; BAUMAN, 1999e HU et al, 2009) e α-caseína (ROSE et al, 2002). Entre as linhagens de células mamárias bovinas mais antigas estão a HH2A e a BMEC+H com ausência de expressão de caseína ou a BMEUV com baixa expressão, que é regulada pela presença de colágeno (ZAVIZION et al, 1996).…”

“…O desenvolvimento de linhagens imortais é muito importante, pois suas células não sofrem senescência e não alteram sua funcionalidade ao longo das passagens (ROSE et al, 2002). Atualmente existem diferentes linhagens celulares imortais para bovinos (GIBSON et al, 1991;HUYNH;ROBITAILLE;TURNER, 1991;GERMAN;BARASH, 2002;ROSE et al, 2002;ZHAO et al, 2010), ovinos (DÜCHLER et al, 1998e ILAN et al, 1998, caprinos (PANTSCHENKO et al, 2000) e suínos (SUN; LI, CHOU, 2006). Outro fator importante é a fase da lactação em que se encontra a fêmea.…”