Methods in Cell Science

, Volume 22, Issue 1, pp 63–66 | Cite as

Preparation of pure cell cultures by cloning

  • Douglas C. McFarland


Due to the heterogeneous nature of animal organs, primary cell cultures potentially contain more than one type of cell. Interpretation of data arising from studies using these mixed cultures can lead to difficulties, because it may not be possible to ascertain which of the cell types present may have responded to a given treatment. While a number of procedures have been used to enrich the cell population of interest, many scientists have resorted to cloning of cells in order to insure the purity of cell cultures. Three major strategies have been used to produce clones, namely, the dilution technique, cloning ring technique, and robotic cell transfer. Successful cloning is dependent on optimization of attachment substrata, basal media composition, serum source, and growth factor/hormone additions to support proliferation of the cell type at clonal (low) density.

Cell culture Cell isolation Clone Method 


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  1. 1.
    Blanton, Jr. JR, Grant AL, McFarland DC, Robinson JP, Bidwell CA (1999). Isolation of two populations of myoblasts from porcine skeletal muscle. Muscle Nerve 22: 43-50.Google Scholar
  2. 2.
    Feldman JL, Stockdale FE (1991). Skeletal muscle satellite cell diversity: Satellite cells form fibers of different types in cell culture. Devel Biol 143: 320-334.Google Scholar
  3. 3.
    Freshney RI (1987). Cloning and selection of specific cell types. In: Culture of Animal Cells: A Manual of Basic Technique, pp 137-153. New York: Alan R. Liss.Google Scholar
  4. 4.
    Johnson AD, McFarland D, Singh YN, Pesall J, Gilkerson K, Vander Wal L (1999). Isolation and characterization of myogenic satellite cells derived from the muscular dystrophic hamster. FASEB J 13(4), pt. 1:A257.Google Scholar
  5. 5.
    Martin BM (1994). Cell Cloning. In: Tissue Culture Techniques: An introduction pp 143-151. Boston, MA: Birkhauser.Google Scholar
  6. 6.
    McFarland DC, Doumit ME, Minshall (1988). The turkey myogenic satellite cell: Optimization of in vitro proliferation and differentiation. Tissue Cell 20: 899-908.Google Scholar
  7. 7.
    McFarland DC, Gilkerson KK, Pesall JE, Walker JS, Yun Y (1995). Heterogeneity in growth characteristics of satellite cell populations. Cytobios 82: 21-27.Google Scholar
  8. 8.
    McFarland DC, Gilkerson KK, Pesall JE, Ferrin NH, Wellenreiter RH (1997). In vitro characteristics of myogenic satellite cells derived from the pectoralis major and biceps femoris muscles of the chicken. Cytobios 91: 45-52.Google Scholar
  9. 9.
    McFarland DC, Pesall JE, Norberg JM, Dvoracek MA (1991). Proliferation of the turkey myogenic satellite cell in a serum-free medium. Comp Biochem Physiol 99A: 163-167.Google Scholar
  10. 10.
    Minshall RD, McFarland DC, Doumit ME (1990). Interaction of insulin-like growth factor I with turkey satellite cells and satellite cell-derived myotubes. Dom Anim Endocr 7: 413-424.Google Scholar
  11. 11.
    Molnar G, Dodson MV (1993). Satellite cells isolated from sheep skeletal muscle are not homogeneous. Basic Appl Myol 3: 173-180.Google Scholar
  12. 12.
    Schultz E, Heckman-Jones J (1990). Labeling characteristics of satellite cells in vivo. J Cell Biol 111: 34a.Google Scholar
  13. 13.
    Sharpe PT (1988). Methods of cell separation. In Burdon RH and van Knippenberg PH (eds). New York: Elsevier.Google Scholar
  14. 14.
    Yun Y, McFarland DC, Pesall JE, Gilkerson KK, Vander Wal LS, Ferrin NH (1997). Variation in response to growth factor stimuli in satellite cell populations. Comp Biochem Physiol 117A: 463-470.Google Scholar

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© Kluwer Academic Publishers 2000

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  • Douglas C. McFarland

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