Cryopreservation of avian lymphoid cells.

E. A. Stephens, J. M. Sharma

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Conditions were standardized for optimum cryopreservation of avian lymphoid cells. Several factors that influenced cryopreservation were examined. The optimum procedure was as follows: a maximum of 50 x 10(6) cells/ml were suspended in the freezing medium, which contained 5-10% dimethyl sulfoxide (DMSO), and the cell suspension was frozen slowly at a rate of -1 C degree/min. The frozen cells were kept at -196 C. Cryopreserved cels were thawed rapidly in a 37 C water bath until the last ice crystal had thawed. The method of diluting out DMSO from thawed cells was critical, and results were best if dilutions were made at room temperature (20-25C) with diluent prewarmed to room temperature. Dilution was begun by adding 0.1 ml diluent to 1.0 ml freshly thawed cell suspension. Thereafter, diluent was added by doubling the volume after each 1-min interval until 12.7 ml of diluent had been added over 6 min. The recovery of viable cells from cryopreserved cells varied from 51.2% to 98.3% (mean 86.0%) for lymphoblastoid line cells and from 24.8% to 87.2% (mean 50.6%) for spleen cells obtained from normal chickens. Viable cryopreserved cells were reactive in a 4-hr Cr-release cytotoxicity assay and responded vigorously to phytohemagglutinin. The standardized method of freezing and thawing avian lymphoid cells may facilitate preservation of large stocks of standard reference cells with predetermined functions for laboratory studies, particularly those involving in vitro assays of cellular immunity.

Original languageEnglish (US)
Pages (from-to)614-627
Number of pages14
JournalAvian Diseases
Volume25
Issue number3
StatePublished - Jul 1981
Externally publishedYes

Fingerprint

Cryopreservation
cryopreservation
Lymphocytes
cells
dimethyl sulfoxide
cell suspension culture
Dimethyl Sulfoxide
ambient temperature
freezing
Freezing
Suspensions
reference standards
phytohemagglutinin
assays
splenocytes
Temperature
thawing
cell-mediated immunity
Ice
crystals

ASJC Scopus subject areas

  • Cancer Research
  • Immunology
  • veterinary(all)

Cite this

Stephens, E. A., & Sharma, J. M. (1981). Cryopreservation of avian lymphoid cells. Avian Diseases, 25(3), 614-627.

Cryopreservation of avian lymphoid cells. / Stephens, E. A.; Sharma, J. M.

In: Avian Diseases, Vol. 25, No. 3, 07.1981, p. 614-627.

Research output: Contribution to journalArticle

Stephens, EA & Sharma, JM 1981, 'Cryopreservation of avian lymphoid cells.', Avian Diseases, vol. 25, no. 3, pp. 614-627.
Stephens EA, Sharma JM. Cryopreservation of avian lymphoid cells. Avian Diseases. 1981 Jul;25(3):614-627.
Stephens, E. A. ; Sharma, J. M. / Cryopreservation of avian lymphoid cells. In: Avian Diseases. 1981 ; Vol. 25, No. 3. pp. 614-627.
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