Korean J Phys Anthropol.
1998 Jun;11(1):131-138.
In vitro Immunization of Human Tonsilar Lymphocytes Using Slice Culture Sytem
- Affiliations
-
- 1Department of Anatomy, Ewha Womans University, Korea.
- 2Department of Microbiology, Ewha Womans University, Korea.
- 3Department of Otolaryngology, Ewha Womans University, Korea.
- 4Department of Thoracic & Cardiovascular Surgery, Ewha Womans University, Korea.
- 5Department of General Surgery, Ewha Womans University, Korea.
- 6Department of Parasitology, Ewha Womans University, Korea.
- 7Department of Internal Medicine, Ewha Womans University, Korea.
Abstract
- It is difficult to immunize human lymphocytes in vitro by conventional cell culture methods. Activation of lymphocytes requires not only specific antigen stimulation but also delicate cell to cell interaction. If the cellular organization could be maintained in culture system, lymphocytes could be immunized in vitro with higher frequency. For the purpose of in vitro immunization of human lymphocytes, we used slice culture system which could maintain morphological and functional organization. Human tonsils resected from eleven -year old boy were evenly divided into two pieces, and one was cultivated in conventional cell culture and the other in slice culture system. In the former system, tonsilar mononuclear cells, separated by Ficoll -Hypaque density gradient centrifugation, were cultivated in RPMI 1640 supplemented with 10% human type AB serum in the cell density of 5 x10 6 /ml. In the latter, tonsillar tissues were sliced into small pieces of 8 mm 3 , and were cultivated in Waymouth MB 752/1 medium supplemented with 10% Human type AB serum, gassed under 5% CO2 and 95% O2 at 37 C. After stabilized for one hour, each system wasw challenged with 50 microgram/ml of KLH or 100 microgram/ml of LPS. At 3, 6, 12 and 24 hours after antigen challenge, culture supernatants were assayed for the specific antibody by ELISA, and cells or tissues were analyzed for the expression of CD23 by flow cytometry. The result showed that tonsilar B lymphocytes in slice culture system expressed CD23 as early as 3 hours after antigen challenge, while those in cell culture system expressed CD23 from 6 hours after challenge. Specific antibodies were detected only in supernatants of slice culture system from 6 hours after challenge. These results suggested thathuman lymphocytes could be immunized in vitro if the cellular organization was maintained.
Keyword
- Full Text Links
-
- Actions
-
Cited
- CITED
-
- Close
- Share
-
- Similar articles
-
- Antibody Response of Mouse Splenocytes Using Mixture of Supernatants of Thymocytes, Adherent and Non-adherent Splenocytes: In-vitro Immunization-II
- The effect of conditioned media cultured from the thymocytes and the sphenocytes on the regulation of antibody isotypes; in vitro immunization-ill
- In vitro immunization-IV: antibody response of mouse splenocytes grown in a mixture of conditioned media, thymocytes, and bone marrow cells
- Organotypic slice culture of the hypothalamic paraventricular nucleus of rat
- Neurotoxic Effect of beta-Amyloid Peptide in Hippocampal Slice Culture
