Isolation of Single Cells, Plant Organs, Culture tissue, Callus, Fresh medium, Suspension culture, Agitation, Cell Clumps

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Home >> Plant Biotechnology and Genomics >> Culture Media and Cell Culture >>Isolation of Single Cells

Isolation of single cells
Single cells can be isolated either from cultured tissues or from intact plant organs, the former being more convenient than the latter. When isolated from cultured tissue, the latter is obtained by culturing an organised tissue (explant) into callus (callus is undifferentiated growth in solid form). The callus may be separated from explant and transferred to fresh medium to get more tissue. Pieces of undifferentiated calli are transferred to liquid medium, which is continuously agitated to obtain a suspension culture.

Agitation of pieces breaks them into smaller clumps and single cells, and also maintains uniform distribution of cell and cell clumps in the medium. It also allows gaseous exchange

Suspension cultures with dissociated single cells can also be obtained from intact plant organs either mechanically (grinding the tissue followed by cleaning, filtration and centrifugation) or enzymatically (treating excised and peeled leaves with macerozyme). The details of technique are available in an advanced book

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