Isolation and Expansion of Mesenchymal Stem/Stromal Cells Derived from Human Placenta Tissue

Pelekanos, Rebecca A.; Sardesai, Varda S.; Futrega, Kathryn; Lott, William B.; Kühn, Michael; Doran, Michael

doi:10.3791/54204

Cited by 66 publications

(64 citation statements)

References 36 publications

(61 reference statements)

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“…As mentioned above, it was puzzling as to why the term placenta, a largely fetal tissue, was reported in the literature to produce MSC cultures purely of maternal origin, even in our own hands . Therefore, a systematic review was carried out in order to ( i) clarify published results on the origin of MSC obtained from the human chorion, ( ii) determine if common methodologies resulted in MSC of a particular origin, and ( iii) summarize the techniques used determine the origin of MSC obtained.…”

Section: Discussionmentioning

confidence: 99%

“…Third trimester placental MSCs were isolated as described previously . A video of the dissection and isolation process, as well as a detailed written protocol, is provided in the following reference .…”

Section: Methodsmentioning

confidence: 99%

“…MSC were isolated as described but with modifications to select placental tissue based on anatomical regions of the villous tissue, specifically the chorionic plate (CP), chorionic villi (CV), or decidua (D) .…”

Section: Methodsmentioning

confidence: 99%

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Avoidance of Maternal Cell Contamination and Overgrowth in Isolating Fetal Chorionic Villi Mesenchymal Stem Cells from Human Term Placenta

Sardesai

Shafiee

Fisk

et al. 2017

Stem Cells Translational Medicine

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Human placenta is rich in mesenchymal stem/stromal cells (MSC), with their origin widely presumed fetal. Cultured placental MSCs are confounded by a high frequency of maternal cell contamination. Our recent systematic review concluded that only a small minority of placental MSC publications report fetal/maternal origin, and failed to discern a specific methodology for isolation of fetal MSC from term villi. We determined isolation conditions to yield fetal and separately maternal MSC during ex vivo expansion from human term placenta. MSCs were isolated via a range of methods in combination; selection from various chorionic regions, different commercial media, mononuclear cell digest and/or explant culture. Fetal and maternal cell identities were quantitated in gender‐discordant pregnancies by XY chromosome fluorescence in situ hybridization. We first demonstrated reproducible maternal cell contamination in MSC cultures from all chorionic anatomical locations tested. Cultures in standard media rapidly became composed entirely of maternal cells despite isolation from fetal villi. To isolate pure fetal cells, we validated a novel isolation procedure comprising focal dissection from the cotyledonary core, collagenase/dispase digestion and explant culture in endothelial growth media that selected, and provided a proliferative environment, for fetal MSC. Comparison of MSC populations within the same placenta confirmed fetal to be smaller, more osteogenic and proliferative than maternal MSC. We conclude that in standard media, fetal chorionic villi‐derived MSC (CV‐MSC) do not grow readily, whereas maternal MSC proliferate to result in maternal overgrowth during culture. Instead, fetal CV‐MSCs require isolation under specific conditions, which has implications for clinical trials using placental MSC. Stem Cells Translational Medicine 2017;6:1070–1084

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Avoidance of Maternal Cell Contamination and Overgrowth in Isolating Fetal Chorionic Villi Mesenchymal Stem Cells from Human Term Placenta

Sardesai

Shafiee

Fisk

et al. 2017

Stem Cells Translational Medicine

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“…Mesenchymal stromal cells (MSCs) are multipotent progenitors derived from different sources, including bone marrow (BM) , adipose tissue ( Van Pham et al, 2014), umbilical cord (UC) blood (Sibov et al, 2012), UC (Van Pham et al, 2016), Wharton's jelly (Ducret et al, 2016), and other tissues (Ducret et al, 2016;Pelekanos et al, 2016;Sharpe, 2016). Although, bone marrow-derived MSCs (BMMSCs) are a popular source of MSCs for research and clinical applications, there are now alternative sources of MSCs, including adipose tissue-derived stem cells (ADSCs) and umbilical cord-derived mesenchymal stem cells (UCMSCs).…”

Section: Introductionmentioning

confidence: 99%

Umbilical cord-derived stem cells (ModulatistTM) show strong immunomodulation capacity compared to adipose tissue-derived or bone marrow-derived mesenchymal stem cells

Pham¹,

Vu²,

Phan³

2016

Biomed Res Ther

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Abstract-Introduction:Mesenchymal stem cells (MSCs) show great promise in regenerative medicine. Clinical applications of MSCs have recently increased significantly, especially for immune diseases. Autologous transplantation is considered a safe therapy. However, its main disadvantages are poor stability and quality of MSCs from patient to patient, and labor-intensive and time-consuming culture procedures. Therefore, allogeneic MSC transplantation has recently emerged as a potential replacement for autologous transplantation. "Off the shelf" MSC products, or socalled "stem cell drugs", have rapidly developed; these products have already been approved in various countries, including Canada, Korea and Japan. This study aims to evaluate a new stem cell product or "drug", termed Modulatist TM , derived from umbilical cord mesenchymal stem cells (UCMSCs), which have strong immunomodulatory properties, compared to bone marrow-derived MSCs (BMMSCs) or adipose tissue-derived stem cells (ADSCs). Methods: Modulatist TM was produced from MSCs derived from whole umbilical cord (UC) tissue (which includes Wharton's jelly and UC), according to GMP compliant procedures. Bone marrow-and adipose tissue-derived MSCs were isolated and proliferated in standard conditions, according to GMP compliant procedures. Immunomodulation mediated by MSCs was assessed by allogenic T cell suppression and cytokine release; role of prostaglandin E2 in the immunomodulation was also evaluated. Results: The results showed that Modulatist TM exhibited stronger immunomodulation than BMMSC and ADSC in vitro. Modulatist TM strongly suppressed allogeneic T cells proliferation and decreased cytokine production, compared to BMMSCs and ADSCs. Conclusion: Modulatist TM is a strong immunomodulator and promising MSC product. It may be useful to modulate or treat autoimmune diseases.

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“…AMSCs were isolated as previously described . Briefly, the amniotic membrane was digested in a rotary incubator at 37°C with a solution of 0.25% trypsin‐EDTA for 30 minutes and then with collagenase type IV solution (Gibco, Carlsbad, California) in complete medium comprising Dulbecco's modified Eagle's medium (DMEM)/F12, 10% fetal bovine serum (FBS) and 1% antibiotic/antimycotic solution, for 1.5 hours.…”

Section: Methodsmentioning

confidence: 99%

Gestational diabetes impacts fetal precursor cell responses with potential consequences for offspring

Algaba-Chueca

Maymó-Masip

Ejarque

et al. 2019

Stem Cells Translational Medicine

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Fetal programming has been proposed as a key mechanism underlying the association between intrauterine exposure to maternal diabetes and negative health outcomes in offspring. To determine whether gestational diabetes mellitus (GDM) might leave an imprint in fetal precursors of the amniotic membrane and whether it might be related to adverse outcomes in offspring, a prospective case‐control study was conducted, in which amniotic mesenchymal stem cells (AMSCs) and resident macrophages were isolated from pregnant patients, with either GDM or normal glucose tolerance, scheduled for cesarean section. After characterization, functional characteristics of AMSCs were analyzed and correlated with anthropometrical and clinical variables from both mother and offspring. GDM‐derived AMSCs displayed an impaired proliferation and osteogenic potential when compared with control cells, accompanied by superior invasive and chemotactic capacity. The expression of genes involved in the inflammatory response (TNFα, MCP‐1, CD40, and CTSS) was upregulated in GDM‐derived AMSCs, whereas anti‐inflammatory IL‐33 was downregulated. Macrophages isolated from the amniotic membrane of GDM mothers consistently showed higher expression of MCP‐1 as well. In vitro studies in which AMSCs from healthy control women were exposed to hyperglycemia, hyperinsulinemia, and palmitic acid confirmed these results. Finally, genes involved in the inflammatory response were associated with maternal insulin sensitivity and prepregnancy body mass index, as well as with fetal metabolic parameters. These results suggest that the GDM environment could program stem cells and subsequently favor metabolic dysfunction later in life. Fetal adaptive programming in the setting of GDM might have a direct negative impact on insulin resistance of offspring.

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Isolation and Expansion of Mesenchymal Stem/Stromal Cells Derived from Human Placenta Tissue

Cited by 66 publications

References 36 publications

Avoidance of Maternal Cell Contamination and Overgrowth in Isolating Fetal Chorionic Villi Mesenchymal Stem Cells from Human Term Placenta

Avoidance of Maternal Cell Contamination and Overgrowth in Isolating Fetal Chorionic Villi Mesenchymal Stem Cells from Human Term Placenta

Umbilical cord-derived stem cells (ModulatistTM) show strong immunomodulation capacity compared to adipose tissue-derived or bone marrow-derived mesenchymal stem cells

Gestational diabetes impacts fetal precursor cell responses with potential consequences for offspring

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