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. A tissue culture is the growth of cells (tissue) separate from the organism. ...

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

Culture of Embryonic organs: Embryonic organ culture is easier than to normal organ from adult animals. Following are the three techniques used for embryo culture.

Organ culture on Plasma clots: It involves the following steps: 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 Petridish; 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.

Organ Culture on Agar

Media solidified with agar are also used for organ culture and these media consist of 7 parts 1% agar in BSS, 3 parts chick embro extract and 3 parts of horse serum. Defined media with or without serum 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 this medium, bur from adult will not survive.

The culture of adult organs or parts from adult animal is more difficult due to their greater requirement of O2. A variety of adult organs (eg., 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 use the use of 95 % oxygen.

DR. K. Choudhary

Whole Embryo Culture:

Culture of ChicK Embryo (Spratt,1956) In this techniquw, 40 hours old embryos were used and the embryo development could be followed for another 24-48 hours, in vitro before the embryo dies.

Steps of Chick Embryo Culture: 1. Prepare a suitable defined media and added to sterile watch glasses, palced on moist absorbent cotton wool pads in petri dish as grown in embryo culture. 2. Incubate the hens egg at 380C for 40-42 hours to provide about a dozen embryos. 3. The shell is wiped with alcohol and broken into sterile evoparating dish containing 50 ml chick saline or BSS. 4. A circular cut is made using scissor into the vitelline membrane around the blastoderm and the latter is transferred to a petri dish containing BSS. 5. The adherent vitelline membrane is removed with the aid of forceps and the embryo is examined under the microscope to determine the stage of development. 6. The blastoderm is transferred to the top of the medium in the watch glass prepared as above. 7. The blstoderm is spread on agar gel (ventral side down) and the excess BSS is removed. 8. Culture is incubated at 37.50C.