Human SAA1-derived amyloid deposition in cell culture: A consistent model utilizing human peripheral blood mononuclear cells and serum-free medium

Wataru Ishii, Juris J. Liepnieks, Toshiyuki Yamada, Merrill Benson, Barbara Kluve-Beckerman

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Amyloid A (AA) amyloidosis is a fatal disease caused by extracellular deposition of fibrils derived from serum AA (SAA). AA amyloid fibril formation has previously been modeled in macrophage cultures using highly amyloidogenic mouse SAA1.1, but attempts to do the same with human SAA invariably failed. Our objective was to define conditions that support human SAA-derived amyloid formation in peripheral blood mononuclear cell (PBMC) cultures. Two conditions were found to be critical-omission of fetal calf serum and use of StemPro34, a lipid-enriched medium formulated for hematopoietic progenitor cells. Cultures maintained in serum-free StemPro34 and provided with recombinant human SAA1 in the complete absence of amyloid-enhancing factor exhibited amyloid deposition within 7 d. Amyloid co-localized with cell clusters that characteristically included cells of fibrocytic/dendritic morphology as well as macrophages. These cells formed networks that appeared to serve as scaffolding within and upon which amyloid accumulated. Cells in amyloid-forming cultures demonstrated increased adherence, survival and expression of extracellular matrix components. Of the three human SAA1 isoforms, SAA1.3 showed the most extensive amyloid deposition, consistent with it being the most prevalent isoform in Japanese patients with AA amyloidosis. Attesting to the reproducibility and general applicability of this model, amyloid formation has been documented in cultures established from eight PBMC donors.

Original languageEnglish
Pages (from-to)61-71
Number of pages11
JournalAmyloid
Volume20
Issue number2
DOIs
StatePublished - Jun 2013

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Serum-Free Culture Media
Amyloid
Blood Cells
Cell Culture Techniques
Serum
Amyloidosis
Protein Isoforms
Macrophages
Serum Amyloid A Protein
Hematopoietic Stem Cells
Dendritic Cells
Extracellular Matrix
Tissue Donors
Lipids

Keywords

  • CCL2
  • Collagen
  • Fetal calf serum
  • Fibrocytes
  • Macrophages
  • Monocytes
  • Serum amyloid A

ASJC Scopus subject areas

  • Internal Medicine

Cite this

Human SAA1-derived amyloid deposition in cell culture : A consistent model utilizing human peripheral blood mononuclear cells and serum-free medium. / Ishii, Wataru; Liepnieks, Juris J.; Yamada, Toshiyuki; Benson, Merrill; Kluve-Beckerman, Barbara.

In: Amyloid, Vol. 20, No. 2, 06.2013, p. 61-71.

Research output: Contribution to journalArticle

Ishii, Wataru ; Liepnieks, Juris J. ; Yamada, Toshiyuki ; Benson, Merrill ; Kluve-Beckerman, Barbara. / Human SAA1-derived amyloid deposition in cell culture : A consistent model utilizing human peripheral blood mononuclear cells and serum-free medium. In: Amyloid. 2013 ; Vol. 20, No. 2. pp. 61-71.
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