A novel cell array technique for high-throughput, cell-based analysis

Reports

Summary

Microarray technology has burgeoned over the past few years from nucleic acid-based arrays to tissue microarrays (TMAs). This study aimed to develop a technique to incorporate cell lines into an array and to demonstrate the usefulness of this technique by performing immunohistochemistry for β-catenin. Cell suspensions were prepared from 23 tumor cell lines. These were fixed in formalin, suspended in agar, and embedded in paraffin to produce a cell block. A “tissue microarrayer” was used to remove triplicate, 0.6 mm-cores from each cell block and to transfer these into a recipient paraffin block at precise coordinates. Immunohistochemistry was used to identify cell lines positive for β-catenin. Cultured cells were successfully incorporated into the microarray, with preservation of cell architecture and even distribution of cells within each core. A total of 18 of 69 cores (26%) were lost in processing. A total of 16 of 23 cell lines were identified as positive for membrane and cytoplasmic β-catenin, and 6 of 23 were negative. Only one cell line was unscorable because of complete core loss. We have developed a “cell microarray” technique for analyzing antigen expression by immunohistochemistry in multiple cell lines in a single expriment. This novel application of microarrays permits high-throughput, cost-efficient analysis, with the potential to rapidly identify markers with potential diagnostic and therapeutic implications in human disease.

Key words

cell lines microarray immunohistochemistry 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. DeRisi, J.; Penland, L.; Brown, P. O., et al. Use of a cDNA microarray to analyse gene expression patterns in human cancer. Nat. Genet. 14:457–460; 1996.PubMedCrossRefGoogle Scholar
  2. Duggan, D. J.; Bittner, M.; Chen, Y.; Meltzer, P.; Trent, J. M. Expression profiling using cDNA microarrays. Nat. Genet. 21:10–14; 1999.PubMedCrossRefGoogle Scholar
  3. Ellison, G.; Klinowska, T.; Westwood, R. F. R.; Docter, E.; French, T.; Fox, J. C. Further evidence to support the melanocytic origin of DDA-MB-435. J. Clin. Pathol. 55:294–299; 2003.Google Scholar
  4. Ginestier, C.; Charafe-Jauffret, E.; Bertucci, F., et al. Distinct and complementary information provided by use of tissue and DNA microarrays in the study of breast tumour markers. Am. J. Pathol. 161:1223–1233; 2002.PubMedGoogle Scholar
  5. Kononen, J.; Bubendorf, L.; Kallioniemi, A., et al. Tissue microarrays for high-throughput molecular profiling of tumour specimens. Nat. Med. 4:844–847; 1998.PubMedCrossRefGoogle Scholar
  6. Masters, J. R.; Thomson, J. A.; Daly-Burns, B., et al. Shor tandem repeat profiling provides an international reference standard for human cell lines. Proc. Natl. Acad. Sci. USA 98:8012–8017; 2001.PubMedCrossRefGoogle Scholar
  7. Mucci, N. R.; Akdas, G.; Manely, S.; Rubin, M. A. Neuroendocrine expression in metastatic prostate cancer: evaluation of high throughput tissue microarrays to detect homogenous protein expression. Hum. Pathol. 31:406–414; 2000.PubMedCrossRefGoogle Scholar
  8. Oode, K.; Furuya, T.; Harada, K.; Kawauchi, S.; Yamamoto, K.; Hirano, T.; Sasaki, K.: The development of a cell array and its combination with laser scanning cytometry allows a high-throughput analysis of nuclear DNA content. Am. J. Pathol. 157:723–728; 2000.PubMedGoogle Scholar
  9. Richter, J.; Wagner, U.; Kononen, J., et al. Tissue microarrays for gene amplification surveys in many different tumour types. Clin. Cancer Res. 5:1966–1975; 1999.Google Scholar
  10. Stephan, J. P.; Schanz, S.; Wong, A.; Schow, P.; Wong, W. L. T. Development of a frozen cell array as a high-throughput approach for cell based analysis. Am. J. Pathol. 161:787–797; 2002.PubMedGoogle Scholar

Copyright information

© Society for In Vitro Biology 2005

Authors and Affiliations

  1. 1.Molecular Medicine UnitUniversity of LeedsLeedsU.K.
  2. 2.Academic Unit of PathologyUniversity of LeedsLeedsU.K.

Personalised recommendations