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. 1973 Oct;15(2):289–302.

Human mixed lymphocyte cultures. Evaluation of microculture technique utilizing the multiple automated sample harvester (MASH)

G B Thurman, D M Strong, A Ahmed, S S Green, K W Sell, R J Hartzman, F H Bach
PMCID: PMC1553877  PMID: 4271568

Abstract

Use of lymphocyte cultures for in vitro studies such as pretransplant histocompatibility testing has established the need for standardization of this technique. A microculture technique has been developed that has facilitated the culturing of lymphocytes and increased the quantity of cultures feasible, while lowering the variation between replicate samples. Cultures were prepared for determination of tritiated thymidine incorporation using a Multiple Automated Sample Harvester (MASH). Using this system, the parameters that influence the in vitro responsiveness of human lymphocytes to allogeneic lymphocytes have been investigated.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BACH F., HIRSCHHORN K. LYMPHOCYTE INTERACTION: A POTENTIAL HISTOCOMPATIBILITY TEST IN VITRO. Science. 1964 Feb 21;143(3608):813–814. doi: 10.1126/science.143.3608.813. [DOI] [PubMed] [Google Scholar]
  2. BAIN B., VAS M. R., LOWENSTEIN L. THE DEVELOPMENT OF LARGE IMMATURE MONONUCLEAR CELLS IN MIXED LEUKOCYTE CULTURES. Blood. 1964 Jan;23:108–116. [PubMed] [Google Scholar]
  3. Bach F. H., Bach M. L., Hartzman R. J. Matching-typing complementarity in histocompatibility testing. Transplant Proc. 1971 Jun;3(2):1099–1104. [PubMed] [Google Scholar]
  4. Bain B. Tritiated-thymidine uptake in mixed leucocyte cultures: effect of specific activity and exposure time. Clin Exp Immunol. 1970 Feb;6(2):255–262. [PMC free article] [PubMed] [Google Scholar]
  5. Bouroncle B. A., Malspeis L., Aschenbrand J. F. Factors affecting the quantitative evaluation of the uni- and bidirectional reactions of mixed leukocyte cultures. Am J Clin Pathol. 1972 Jul;58(1):17–25. doi: 10.1093/ajcp/58.1.17. [DOI] [PubMed] [Google Scholar]
  6. Brody J. A., Huntley B. Human lymphocytes cultured in microplates. Nature. 1965 Dec 18;208(5016):1232–1233. doi: 10.1038/2081232b0. [DOI] [PubMed] [Google Scholar]
  7. Caspary E. A., Hughes D. Liquid scintillation counting techniques in lymphocyte transformation in vitro measured by tritiated thymine uptake. J Immunol Methods. 1972 May;1(3):263–272. doi: 10.1016/0022-1759(72)90004-x. [DOI] [PubMed] [Google Scholar]
  8. Darzynkiewicz Z., Jacobson B. HEPES-buffered media in lymphocyte cultures. Proc Soc Exp Biol Med. 1971 Feb;136(2):387–393. doi: 10.3181/00379727-136-35271. [DOI] [PubMed] [Google Scholar]
  9. Eagle H. Buffer combinations for mammalian cell culture. Science. 1971 Oct 29;174(4008):500–503. doi: 10.1126/science.174.4008.500. [DOI] [PubMed] [Google Scholar]
  10. Green S. S., Sell K. W. Mixed leukocyte stimulation of normal peripheral leukocytes by autologous lymphoblastoid cells. Science. 1970 Nov 27;170(3961):989–990. doi: 10.1126/science.170.3961.989. [DOI] [PubMed] [Google Scholar]
  11. Halpern B., Amache N. Diagnosis of drug allergy in vitro with the lymphocyte transformation test. J Allergy. 1967 Sep;40(3):168–181. doi: 10.1016/0021-8707(67)90006-8. [DOI] [PubMed] [Google Scholar]
  12. Han T., Sokal J. E. Lymphocyte response to phytohemagglutinin in Hodgkin's disease. Am J Med. 1970 Jun;48(6):728–734. doi: 10.1016/s0002-9343(70)80008-0. [DOI] [PubMed] [Google Scholar]
  13. Hartzman R. J., Bach M. L., Bach F. H., Thurman G. B., Sell K. W. Precipitation of radioactively labeled samples: a semi-automatic multiple-sample processor. Cell Immunol. 1972 Jun;4(2):182–186. doi: 10.1016/0008-8749(72)90018-4. [DOI] [PubMed] [Google Scholar]
  14. Hartzman R. J., Segall M., Bach M. L., Bach F. H. Histocompatibility matching. VI. Miniaturization of the mixed leukocyte culture test: a preliminary report. Transplantation. 1971 Mar;11(3):268–273. doi: 10.1097/00007890-197103000-00005. [DOI] [PubMed] [Google Scholar]
  15. Jackson S. M., Garrett J. V., Craig A. W. Lymphocyte transformation changes during the clinical course of Hodgkin's disease. Cancer. 1970 Apr;25(4):843–850. doi: 10.1002/1097-0142(197004)25:4<843::aid-cncr2820250413>3.0.co;2-1. [DOI] [PubMed] [Google Scholar]
  16. Nagel G. A., Holland J. F. The mixed lymphocyte tumor cell culture and the fluorescein diacetate cytotoxicity test. Two useful methods for the demonstration of cell mediated tumor specific immunity in man. Acta Haematol. 1970;44(3):129–141. doi: 10.1159/000208671. [DOI] [PubMed] [Google Scholar]
  17. Parker J. W., Lukes R. J. A microculture method for lymphocyte transformation studies in the clinical laboratory. Am J Clin Pathol. 1971 Aug;56(2):174–180. doi: 10.1093/ajcp/56.2.174. [DOI] [PubMed] [Google Scholar]
  18. Polet H. The mixed lymphocyte culture as a tool for the quantitative determination of HL-A differences. Transplantation. 1972 May;13(5):472–480. doi: 10.1097/00007890-197205000-00005. [DOI] [PubMed] [Google Scholar]
  19. Rádl J., Dooren L. J., Eijsvoogel V. P., van Went J. J., Hijmans W. An immunological study during post-transplantation follow-up of a case of severe combined immunodeficiency. Clin Exp Immunol. 1972 Feb;10(2):367–382. [PMC free article] [PubMed] [Google Scholar]
  20. Sample W. F., Chretien P. B. Thymidine kinetics in human lymphocyte transformation: determination of optimal labelling conditions. Clin Exp Immunol. 1971 Sep;9(3):419–427. [PMC free article] [PubMed] [Google Scholar]
  21. Shons A. R., Etheredge E. E., Najarian J. S. Variables in the quantitation of the mixed leucocyte response by tritiated thymidine uptake. Clin Exp Immunol. 1972 Nov;12(3):351–356. [PMC free article] [PubMed] [Google Scholar]
  22. Steele R. W., Limas C., Thurman G. B., Schuelein M., Bauer H., Bellanti J. A. Familial thymic aplasia. Attempted reconstitution with fetal thymus in a Millipore diffusion chamber. N Engl J Med. 1972 Oct 19;287(16):787–791. doi: 10.1056/NEJM197210192871602. [DOI] [PubMed] [Google Scholar]

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