Isolation of human multipotent mesenchymal stem cells from second-trimester amniotic fluid using a novel two-stage culture protocol

Tsai, Ming‐Song; Lee, Jia-Ling; Chang, Yu-Jen; Hwang, Shiaw-Min

doi:10.1093/humrep/deh279

Cited by 595 publications

(507 citation statements)

References 25 publications

Supporting

Mentioning

446

Contrasting

Unclassified

Order By: Relevance

“…After these treatments, AFCs gave rise to adipocytes and osteoblasts, when fibroblasts were the predominant cell population. In this regard, it has been reported [10,12,13] that amniotic fluid mesenchymal stem cells display a multilineage differentiation potential into fibroblasts, adipocytes and osteocytes. We suggest that the limited differentiation potential towards mesenchymal lineages that we found in our AFC cultures could be due to the cell heterogeneity with high percentage of epithelial cells.…”

Section: Discussionmentioning

confidence: 99%

“…Amniotic epithelial cells obtained from cesarean sections have been shown to express markers for neuronal, glial and progenitor cells and to differentiate into neuron-like cells in the ischemic brain of adult rats [7][8][9]. Moreover, there is evidence that amniotic fluid contains fetal mesenchymal stem cells with a multilineage differentiation potential [10][11][12][13]. In this study we extensively characterized unselected amniotic cells and tested their multilineage differentiation capacity in vitro.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Molecular and phenotypic characterization of human amniotic fluid cells and their differentiation potential

et al. 2006

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Molecular and phenotypic characterization of human amniotic fluid cells and their differentiation potential

et al. 2006

View full text Add to dashboard Cite

“…In recent years, several studies have demonstrated the ability to isolate amniotic membrane stem cells (AMSCs) and amniotic fluid stem cells (AFSCs) (In 't Anker et al 2003;Tsai et al 2004;Tsai et al 2006). These cells have phenotypes attributable to pluripotent cells and can differentiate into different cell types due to the three different embryonic layers (De Coppi et al 2007).…”

Section: Introductionmentioning

confidence: 99%

Isolation, characterization, and in vitro differentiation of ovine amniotic stem cells

et al. 2010

View full text Add to dashboard Cite

Stem cell (SC) regenerative therapy represents an emerging strategy for the treatment of human diseases. Since amniotic fluid-derived cells have been recently proposed as a promising source of human SCs, the present research aimed to amplify in vitro and characterize ovine amniotic fluid-derived SCs collected from the membranes (AMSCs) or fluid (AFSCs). These cells were found to proliferate, express the pluripotent SC markers OCT-4 and TERT, and differentiate in both osteogenic and smooth muscle lineages in vitro. However, AMSCs presented an earlier down-regulation of SC markers and a faster rate of differentiation. Thus, AMSCs and AFSCs may represent sources of characterized pluripotent SCs that can be easily collected and amplified in vitro. These ovine SCs may be used in preclinical studies on large animals to develop future human therapies.

show abstract

“…In addition to the deliberately simple and easily reproducible methodology for isolation of amniotic and placental MSCs described here and in our previous work, other methods that rely on oxygen tension, two-stage cultures, or alternative media formulations have been reported [11,12]. It remains to be seen whether these alternatives would lead to any advantage, or disadvantage, in clinical-grade processing.…”

Section: Discussionmentioning

confidence: 96%

Preclinical regulatory validation of a 3-stage amniotic mesenchymal stem cell manufacturing protocol

Steigman

Armant

Bayer-Zwirello

et al. 2008

Journal of Pediatric Surgery

View full text Add to dashboard Cite

Purpose-Due to the 4-6-month interval between a diagnostic amniocentesis and birth, clinical application of amniotic mesenchymal stem cell (AMSC)-based therapies demands a 3-stage cell manufacturing process, including isolation/primary expansion, cryopreservation, and thawing/ secondary expansion. We sought to determine the feasibility and cell yield of such a staged cell manufacturing process, within regulatory guidelines. Methods-HumanAMSCs isolated from diagnostic amniocentesis samples (n=11) were processed under FDA-accredited Good Manufacturing Practice. Expanded cells were characterized by flow cytometry and cryopreserved for 3-5 months. Cell release criteria included: >90% CD29+, CD73+, and CD44+; <5% CD34+ and CD45+; negative mycoplasma QPCR and endotoxin assay; and ≥70% viability.Results-Isolation and ample expansion of AMSCs was achieved in 54.5% (6/11) of the samples. Early processing and at least a 2mL sample were necessary for reliable cell manufacturing. Cell yield before cryopreservation was 223.2±65.4×10 6 cells (44.6-fold expansion), plus a 14.7×10 6 -cell backup, after 36.3±7.8 days. Cell viability post-thaw was 88%. Expanded cells maintained a multipotent mesenchymal progenitor profile.Conclusions-Human amniotic mesenchymal stem cells can be manufactured in large numbers from diagnostic amniocentesis, by an accredited staged processing, under definite procurement guidelines. These data further support the viability of clinical trials of amniotic mesenchymal stem cell-based therapies.

show abstract

Isolation of human multipotent mesenchymal stem cells from second-trimester amniotic fluid using a novel two-stage culture protocol

Cited by 595 publications

References 25 publications

Molecular and phenotypic characterization of human amniotic fluid cells and their differentiation potential

Molecular and phenotypic characterization of human amniotic fluid cells and their differentiation potential

Isolation, characterization, and in vitro differentiation of ovine amniotic stem cells

Preclinical regulatory validation of a 3-stage amniotic mesenchymal stem cell manufacturing protocol

Contact Info

Product

Resources

About