Advertisement

Urological Research

, Volume 21, Issue 1, pp 49–53 | Cite as

A method for isolating smooth muscle cells from pig urinary bladder with low concentrations of collagenase and papain: the relation between calcium concentration and isolated cell length

  • R. Schot
  • E. van Asselt
  • R. van Mastrigt
Original Articles

Summary

The present study describes a method for isolating single smooth muscle cells from pig urinary bladder using a continuous resuspension device. Low concentrations of collagenase and papain were sufficient to obtain a high yield of viable smooth muscle cells, which remained viable for about 3–4 h as tested with fluorescein diacetate. Addition of fetal calf serum increased the lifespan of the isolated cells and the percentage of contractile smooth muscle cells, but caused spontaneous shortening of the cells. The length and volume of the isolated smooth muscle cells depended on the calcium concentration used in the isolation buffer solution. The isolated muscle cells were apparently relaxed if a calcium concentration less than 1.0 mmol/l was used in the isolation medium. In higher calcium concentrations the isolated cells were significantly shorter, probably as a result of a contraction caused by mechanical stimulation of the cells during the isolation procedure.

Key words

Bladder Calcium Cell isolation Contration Smooth muscle 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Asselt E van, Schot R, Mastrigt R van (1991) Cell length measurements in longitudinal smooth muscle strips of the pig urinary bladder. Neurourol Urodyn 10:327Google Scholar
  2. 2.
    Bagby RM, Young AM, Fisher BA, McKinnon M (1971) Contraction of single smooth muscle cells from Bufo marinus stomach. Nature 234:351Google Scholar
  3. 3.
    Bülbring E, Brading AF, Jones, Tomita T (1981) Structure of smooth muscles. In: Gabella G (ed) Smooth muscle. Edward Arnold, London, p 38Google Scholar
  4. 4.
    Douglas WR (1972) Of pigs and men and research. Space Life Sci 3:226Google Scholar
  5. 5.
    Dijk AM van, Laird JD (1984) Characterization of single isolated vascular smooth muscle cells from bovine coronary artery. Blood Vessels 21:267Google Scholar
  6. 6.
    Driska SP, Porter R (1986) Isolation of smooth muscle cells from swine carotid artery by digestion with papain. Am J Physiol 251:C474Google Scholar
  7. 7.
    Fay FS, Delise CM (1973) Contraction of isolated smooth muscle cells: structural changes. Proc Natl Acad Sci USA 70:641Google Scholar
  8. 8.
    Fisher BA, Bagby RM (1977) Reorientation of myofilaments during contraction of a vertebrate smooth muscle. Am J Physiol 232:C5Google Scholar
  9. 9.
    Glerum JJ, Mastrigt R van, Romijn JC, Griffiths DJ (1987) Isolation and individual electrical stimulation of single smooth-muscle cells from the urinary bladder of the pig. J Muscle Res Cell Motil 8:125Google Scholar
  10. 10.
    Gosling JA, Dixon JS, Humpherson JR (1983) Gross and microscopic anatomy of the urinary bladder. In: Functional anatomy of the urinary tract. Gower Medical Publishing, LondonGoogle Scholar
  11. 11.
    Griffiths DJ, Mastrigt R van, Duyl WA van, Coolsaet BLRA (1979) Active mechanical properties of the smooth muscle of the urinary bladder. Med Biol Eng Comput 17:281Google Scholar
  12. 12.
    Koeveringe GA van, Mastrigt R van (1991) Excitatory pathways in smooth muscle investigated by phaseplot analysis of isometric force development. Am J Physiol 261:R138Google Scholar
  13. 13.
    Maruyama I, Yoshida C, Kobayashi M, Oyamada H, Momose K (1987) Preparation of single smooth muscle cells from guinea-pig taenia coli by combinations of purified collagenase and papain. J Pharmacol Methods 18:151Google Scholar
  14. 14.
    Maruyama I, Yoshida C, Nakayama M, Hasegawa T, Momose K (1988) Improvement of a procedure for preparing single smooth muscle cells rom guinea-pig taenia coli by purified collagenase and papain. J Pharmacol Methods 19:155Google Scholar
  15. 15.
    Mastrigt R van, Glerum JJ (1985) Electrical stimulation of smooth muscle strips from the urinary bladder of the pig. J Biomed Eng 7:2Google Scholar
  16. 16.
    Mastrigt R van, Koopal JWB, Hak J, Wetering J van de (1986) Modelling the contractility of urinary bladder smooth muscle using isometric contractions. Am J Physiol 251:978Google Scholar
  17. 17.
    Schot R, Koeveringe AJ van, Mastrigt R van (1990) An improved procedure for isolating smooth muscle cells from the pig urinary bladder. J Muscle Res Cell Motil 1:86Google Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • R. Schot
    • 1
  • E. van Asselt
    • 1
  • R. van Mastrigt
    • 1
  1. 1.Department of UrologyErasmus University RotterdamRotterdamThe Netherlands

Personalised recommendations