Hematopoietic long-term culture (hLTC) for human cytomegalovirus latency and reactivation

Mahadevaiah Umashankar, Felicia Goodrum

Research output: Contribution to journalArticlepeer-review

30 Scopus citations

Abstract

Of the many research challenges posed by human cytomegalovirus latency, perhaps the most notable is the requirement for primary hematopoietic cell culture. Culturing hematopoietic subpopulations while maintaining physiological relevance must be given utmost consideration. We describe a long-standing primary CD34 + hematopoietic progenitor cell (HPCs) system as an experimental model to study human cytomegalovirus (HCMV) latency and reactivation. Key aspects of our model include infection of primary human CD34 + HPCs prior to ex vivo expansion, maintenance of undifferentiated cells in a long-term culture with a stromal cell support, and an assay to quantitate infectious centers produced prior to and following a reactivation stimulus. Our method offers a unique way to quantitatively assess HCMV latency and reactivation to study the contribution of viral and host genes in latency and reactivation.

Original languageEnglish (US)
Pages (from-to)99-112
Number of pages14
JournalMethods in Molecular Biology
Volume1119
DOIs
StatePublished - 2014
Externally publishedYes

Keywords

  • CD34 + hematopoietic cells
  • Extreme limiting dilution analysis
  • Human cytomegalovirus
  • Latency
  • Reactivation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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