216
Methods for Setting Up Primary Cultures Specific to Animal Groups
Results
In TTP medium, most tissue fragments adhere to the substrate and cell
migration red from the explants. Some cells migrated by changing their
form. The migrated cells form networks by connecting their cytoplasmic processes. Mitosis was not observed frequently. Macrophage-like cells
and epidermal cells with cilia can be seen. The latter cell type showed a
rotating movement due to the action of the cilia.
Comments
Murray (1931) cultured fragments of planarians in simple salt solution
(0.5 g NaCl, 0.01 g KCI, 0.07 g CaCI2·2H 2 0, and 0.01 g NaHC0 3 in 150 ml
H 2 0). The period of cell survival was increased markedly by the addition of the planarian digest into the salt solution. The digest was prepared as follows: 1.3 g living planarians were placed in 33 ml N/20 HCI
plus 0.1 g Fairchild's pepsin, covered with a thin layer of toluol, and incubated at 40°C for 2.5 h. At the end of that time, one volume redistilled
water was added to the mixture, and the pH was adjusted to approximately 7.4 with normal NaOH, and the mixture boiled down to approximately 10 ml. This solution was then added to 200 ml of the simple salt
solution described above. The solution was sterilized by autoclaving.
Ansevin and Buchsbaum (1961) cultured dissociated cells from the whole
body of Dugesia tigrina in a medium consisting of a mixture of a planarian tissue extract and physiological solution. If the medium was solidified with 2% agar, cell migration occurred, and the cells showed regeneration. In addition, morphallaxis occurred inside the explants. In a liquid medium, cell migration was inhibited, but restitutional bodies were
formed and the larger pieces of tissue survived for longer.
Subprotocol 5
Hanging Drop Culture of Explants from P/anaria (Turbellaria)
Murray (1926,1927) cultured explants from various parts of Planaria in
a relatively simple culture medium. The following procedure are her
methods.
Materials
Equipment - Dissecting microscope
- Scissors
- Forceps
- Needles
- Pasteur pipettes
- Maximov slides
- Cover glasses
- Depression slide glasses
Methods for Setting Up Primary Cultures Specific to Animal Groups
Results
In TTP medium, most tissue fragments adhere to the substrate and cell
migration red from the explants. Some cells migrated by changing their
form. The migrated cells form networks by connecting their cytoplasmic processes. Mitosis was not observed frequently. Macrophage-like cells
and epidermal cells with cilia can be seen. The latter cell type showed a
rotating movement due to the action of the cilia.
Comments
Murray (1931) cultured fragments of planarians in simple salt solution
(0.5 g NaCl, 0.01 g KCI, 0.07 g CaCI2·2H 2 0, and 0.01 g NaHC0 3 in 150 ml
H 2 0). The period of cell survival was increased markedly by the addition of the planarian digest into the salt solution. The digest was prepared as follows: 1.3 g living planarians were placed in 33 ml N/20 HCI
plus 0.1 g Fairchild's pepsin, covered with a thin layer of toluol, and incubated at 40°C for 2.5 h. At the end of that time, one volume redistilled
water was added to the mixture, and the pH was adjusted to approximately 7.4 with normal NaOH, and the mixture boiled down to approximately 10 ml. This solution was then added to 200 ml of the simple salt
solution described above. The solution was sterilized by autoclaving.
Ansevin and Buchsbaum (1961) cultured dissociated cells from the whole
body of Dugesia tigrina in a medium consisting of a mixture of a planarian tissue extract and physiological solution. If the medium was solidified with 2% agar, cell migration occurred, and the cells showed regeneration. In addition, morphallaxis occurred inside the explants. In a liquid medium, cell migration was inhibited, but restitutional bodies were
formed and the larger pieces of tissue survived for longer.
Subprotocol 5
Hanging Drop Culture of Explants from P/anaria (Turbellaria)
Murray (1926,1927) cultured explants from various parts of Planaria in
a relatively simple culture medium. The following procedure are her
methods.
Materials
Equipment - Dissecting microscope
- Scissors
- Forceps
- Needles
- Pasteur pipettes
- Maximov slides
- Cover glasses
- Depression slide glasses