Isolation of human monocytes on re-orienting gradients of percoll

John A. Hardin, James T. Downs

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

We have developed a rapid and simple method for isolating monocytes from peripheral blood. After mononuclear cells are isolated on a Ficoll-Hypaque gradient, the monocytes are purified by sedimentation for 5 min through a re-orienting gradient of Percoll. The method is easily reproducible and yields approximately half of the monocytes in starting blood, with minimal contamination by lymphocytes. Although both monocytes and lymphocytes have similar densities, separation occurs because these cells migrate at differing rates through the gradient to reach their equilibration point. The recovered monocytes are viable and functional; over 95% of them exclude trypan blue and they adhere to glass and plastic and ingest latex particles normally. By virtue of the rapidity and gentle nature of its centrifugation steps, the method exposes cells to only very limited non-physiologic conditions. For this reason and because the monocytes are maintained in suspension, opportunities for functional alterations are minimized.

Original languageEnglish (US)
Pages (from-to)1-6
Number of pages6
JournalJournal of Immunological Methods
Volume40
Issue number1
DOIs
StatePublished - Jan 16 1981
Externally publishedYes

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Monocytes
Lymphocytes
Diatrizoate
Ficoll
Trypan Blue
Centrifugation
Microspheres
Plastics
Glass
Percoll
Suspensions

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

Isolation of human monocytes on re-orienting gradients of percoll. / Hardin, John A.; Downs, James T.

In: Journal of Immunological Methods, Vol. 40, No. 1, 16.01.1981, p. 1-6.

Research output: Contribution to journalArticle

Hardin, John A. ; Downs, James T. / Isolation of human monocytes on re-orienting gradients of percoll. In: Journal of Immunological Methods. 1981 ; Vol. 40, No. 1. pp. 1-6.
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