Establishment and characterization of bovine mammary epithelial cell lines

Abstract

One bovine mammary epithelial cell clone, designated PS-BME-Cl, and two bovine mammary epithelial cell lines, designated PS-BME-L6 and PS-BME-L7, were derived from mammary tissue of a pregnant (270 day) Holstein cow. The cells exhibit the distinctive morphologic characteristics of mammary epithelial cells and express the milk fat globule membrane protein, PAS-III. They form domes when cultured on plastic substrata and acinilike aggregates when cultured on a collagen matrix. These cells are capable of synthesizing and secreting alpha-lactalbumin and alpha-s1-casein when cultured on a collagen matrix in the presence of insulin, cortisol, and prolactin. The cells have a near-normal diploid number and do not grow in suspension culture. When transplanted to the cleared mammary fat pads of female athymic nude mice, the cells readily proliferate forming noninvasive palpable spherical cellular masses within 8 wk after inoculation. The cells may become a useful tool to study the regulation of ruminant mammary epithelial cell growth and differentiation.

In Vitro Culture and Characterization of a Mammary Epithelial Cell Line from Chinese Holstein Dairy Cow

“…Compared with previous studies showing a doubling of cell proliferation between 24–36 h of culture [10], [26], the latent phase of the isolated primary epithelial cells in our study was >72 h. A possible reason for this effect is that the cells obtained in our study were not of clonal lineage, as in previous studies [5]–[6]. Growth curves showed that there was no significant difference in proliferation between the cells before and after frozen reservation.…”

“…Collagenase dissociation was used successfully during isolation and culture of bovine epithelial cells in vitro [5]–[6]. Tissue culture is another method which was successfully used to isolate bovine mammary epithelial cells [7].…”

“…Tissue culture is another method which was successfully used to isolate bovine mammary epithelial cells [7]. Bovine mammary epithelial cell lines reported in the past include BMEC+H [8], MAC-T [1], PS-BME [5], BME-UV [9] and L-1 [10]. In these studies, the isolated cells were not immortal, thus, additional work was required to develop a bovine mammary epithelial cell line.…”

β-casein gene expression by in vitro cultured bovine mammary epithelial cells derived from developing mammary glands

“…Several lines of immortalized BMECs have been reported in the literature (Gibson et al, 1991;Huynh et al, 1991;Huynh and Pollak, 1995;Zavizion et al, 1996). These transformed lines do not express milk proteins or express very low levels of protein; this modest expression is commonly independent of some or all factors that control in vivo lactogenesis, such as hormones and extracellular matrix (Huynh et al, 1991).…”

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

“…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.…”