Organ In Vitro Culture: What Have We Learned about Early Kidney Development?

Figure 1

Method setup, from Trowel to Saxen: in 1954 Trowell introduced a new method to culture whole organs. He used a metal grid in support of a cotton sheet or filter that would hold the embryonic kidney; the cotton sheet was soaked with culture medium (see (a)) [2]. Culture medium was added only to the level of the grid to cover the tissue with a thin layer of the medium due to surface tension [2]. This set up became very useful to studying aspects of nutrition and metabolism in vitro (a). One year later in 1956, Grobstein slightly modified the method and introduced the “on-the-cloth” (see (b)) and “supported-ring” (c) methods [22]. Both methods used the embryonic spinal cord (eSC) from mouse [43] as inducer. The noninduced mesenchyme was placed on a filter, and a second filter was used to support the eSC. The layout was later called “sandwich type culture.” The “on-the-cloth” method used glass-cloth as a support for the tissue cultures on the filters (as in (b)) and the “supported-ring” used a Plexiglas ring onto which the filters were cemented (c). In 1962, Saxen combined and simplified these methods (d). He cultured the noninduced mesenchyme and spinal cord separated by a filter [22] on a metal grid [2] to support the tissues on the filter in a simple culture dish (e) [117]. Saxen’s modernization has been well taken by others and it is still successfully used nowadays.

(a)

(b)

(c)

(d)

(e)