High-yield isolation, expansion, and differentiation of murine bone marrow-derived mesenchymal stem cells using fibrin microbeads (FMB)

Rachel Rivkin, Alon Ben-Ari, Ibrahim Kassis, Lior Zangi, Elena Gaberman, Lilia Levdansky, Gerard Marx, Raphael Gorodetsky

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Transplantation of adult mesenchymal stem cells (MSCs) could provide a basis for tissue regeneration. MSCs are typically isolated from bone marrow (BM) based on their preferential adherence to plastic, although with low efficiency in terms of yield and purity. Extensive expansion is needed to reach a significant number of MSCs for any application. Fibrin microbeads (FMB) were designed to attach mesenchymal cells and to provide a matrix for their expansion. The current study was aimed at isolating a high yield of purified BM-derived mouse MSCs based on their preferential adherence and proliferation on FMB in suspension cultures. MSCs could be downloaded to plastics or further expanded on FMB. The yield of MSCs obtained by the FMB isolation technique was about one order of magnitude higher than that achieved by plastic adherence, suggesting that these cells are more abundant than previously reported. FMB-isolated cells were classified as MSCs by their fibroblastic morphology, self-renewal ability, and expression profile of their surface antigens, as examined by flow cytometry and immunostaining. In cell culture, the isolated MSCs could be induced to differentiate into three different mesodermal lineages, as demonstrated by histochemical stains and by RT-PCR analyses of tissue-specific genes. MSCs were also able to differentiate into osteocytes while still cultured on FMB. Our results suggest that FMB might serve as an efficient platform for the isolation, expansion, and differentiation of mouse BM-derived MSCs to be subsequently implanted for tissue regeneration.

Original languageEnglish (US)
Pages (from-to)157-175
Number of pages19
JournalCloning and Stem Cells
Volume9
Issue number2
DOIs
StatePublished - Jul 10 2007
Externally publishedYes

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Fibrin
Mesenchymal Stromal Cells
Microspheres
Bone Marrow
Plastics
Regeneration
Osteocytes
Adult Stem Cells
Surface Antigens
Suspensions
Flow Cytometry
Coloring Agents
Cell Culture Techniques
Transplantation
Polymerase Chain Reaction
Genes

ASJC Scopus subject areas

  • Biotechnology
  • Developmental Biology
  • Applied Microbiology and Biotechnology

Cite this

High-yield isolation, expansion, and differentiation of murine bone marrow-derived mesenchymal stem cells using fibrin microbeads (FMB). / Rivkin, Rachel; Ben-Ari, Alon; Kassis, Ibrahim; Zangi, Lior; Gaberman, Elena; Levdansky, Lilia; Marx, Gerard; Gorodetsky, Raphael.

In: Cloning and Stem Cells, Vol. 9, No. 2, 10.07.2007, p. 157-175.

Research output: Contribution to journalArticle

Rivkin, Rachel ; Ben-Ari, Alon ; Kassis, Ibrahim ; Zangi, Lior ; Gaberman, Elena ; Levdansky, Lilia ; Marx, Gerard ; Gorodetsky, Raphael. / High-yield isolation, expansion, and differentiation of murine bone marrow-derived mesenchymal stem cells using fibrin microbeads (FMB). In: Cloning and Stem Cells. 2007 ; Vol. 9, No. 2. pp. 157-175.
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