Organ culture

Organ culture is a development from tissue culture methods of research, the organ culture is able to accurately model functions of an organ in various states and conditions by the use of the actual in vitro organ itself.

Parts of an organ or a whole organ can be cultured in vitro. The main objective is to maintain the architecture of the tissue and direct it towards normal development. In this technique, it is essential that the tissue is never be disrupted or damaged. It thus requires careful handling. The media used for a growing organ culture are generally the same as those used for tissue culture. The techniques for organ culture can be classified into (i) those employing a solid medium and (ii) those employing liquid medium.

Methodology

Embryonic organ culture is an easier alternative to normal organ culture derived from adult animals. The following are three techniques employed for embryonic organ culture.

Plasma clot method

The following are general steps in organ culture on plasma clots.

1. Prepare a plasma clot by mixing 15 drops of plasma with five drops of embryo extract in a watch glass.
2. Place a watch glass on a pad of cotton wool in a petri dish; cotton wool is kept moist to prevent excessive evaporation from the dish.
3. Place a small, carefully dissected piece of tissue on top of the plasma clots in watch glass.

The technique has now been modified, and a raft of lens paper or rayon net is used on which the tissue is placed. Transfer of the tissue can then be achieved by raft easily. Excessive fluid is removed and the net with the tissue placed again on the fresh pool of medium.Parts of an organ or a whole organ can be cultured in vitro. The main objective is to maintain the architecture of the tissue and direct it towards normal development. In this technique, it is essential that the tissue never be disrupted or damaged. It thus requires careful handling. The media used for a growing organ are generally the same as those used for tissue culture. The techniques for organ culture can be classified into (i) those employing a solid medium and (ii) those employing liquid medium.

Agar gel method

Media solidified with agar are also used for organ culture and these media consist of 7 parts 1% agar in BSS, 3 parts chick embryo extract and 3 parts of horse serum. Defined media with or without serum are also used with agar. The medium with agar provides the mechanical support for organ culture. It does not liquefy. Embryonic organs generally grow well on agar, but adult organ culture will not survive on this medium.

The culture of adult organs or parts from adult animals is more difficult due to their greater requirement of oxygen. A variety of adult organs (e.g. the liver) have been cultured using special media with special apparatus (Towell’s II culture chamber). Since serum was found to be toxic, serum-free media were used, and the special apparatus permitted the use of 95% oxygen.

Raft Methods

In this approach the explant is placed onto a raft of lens paper or rayon acetate, which is floated on serum in a watch glass. Rayon acetate rafts are made to float on the serum by treating their 4 corners with silicone.

Similarly, floatability of lens paper is enhanced by treating it with silicone. On each raft, 4 or more explants are usually placed.

In a combination of raft and clot techniques, the explants are first placed on a suitable raft, which is then kept on a plasma clot. This modification makes media changes easy, and prevents the sinking of explants into liquefied plasma.

Grid Method

Initially devised by Trowell in 1954, the grid method utilizes 25 mm x 25 mm pieces of a suitable wire mesh or perforated stainless steel sheet whose edges are bent to form 4 legs of about 4 mm height.

Skeletal tissues are generally placed directly on the grid but softer tissues like glands or skin are first placed on rafts, which are then kept on the grids.

The grids themselves are placed in a culture chamber filled with fluid medium up to the grid; the chamber is supplied with a mixture of O₂ and CO₂ to meet the high O₂ requirements of adult mammalian organs. A modification of the original grid method is widely used to study the growth and differentiation of adult and embryonic tissues.

Limitations

• Results from organ cultures are often not comparable to those from whole animals studies, e.g. in studies on drug action since the drugs are metabolized in vivo but not in vitro.

Current progress

In April 2006, scientists reported a successful trial of seven bladders grown in-vitro and given to humans.^[1] A bladder has been cultured by Anthony Atala of the Wake Forest Institute for Regenerative Medicine in Winston-Salem, North Carolina. A jawbone has been cultured at Columbia University, a lung has been cultured at Yale. A beating rat heart has been cultured by Doris Taylor at the University of Minnesota. An artificial kidney has been cultured by H. David Humes at the University of Michigan. ^[2]

See also

• Cell culture
• Tissue culture

References

1. ^ "Lab-grown bladder shows big promise", New Scientist, 2006, issue 2546, http://www.newscientist.com/channel/health/mg19025464.200-labgrown-bladder-shows-big-promise.html
2. ^ Allready cultured organs to date (2011)

External links

Organ culture Techniques

• Fetal Thymus Organ Culture