Isolation, culture and identification of epidermal stem cells from newborn mouse skin

Reiisi, Somayeh; Esmaeili, Fariba; Shirazi, Abolfazl

doi:10.1007/s11626-009-9245-y

Cited by 25 publications

(22 citation statements)

References 45 publications

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“…A histological observation also supported the proliferative rate and plasticity of NIH3T3 cells. High chromatin content and frequent appearance of dividing nuclei indicated progenitor-like features, which are in accordance with earlier studies done with pluripotent stem cells [1,5,[38][39][40].…”

Section: In Vivo Teratoma Formation Reconfirms Pluripotency In Nih3t3supporting

confidence: 91%

Basal Expression of Pluripotency-Associated Genes Can Contribute to Stemness Property and Differentiation Potential

Dadheech

Srivastava

Belani

et al. 2013

Stem Cells and Development

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Pluripotency and stemness is believed to be associated with high Oct-3/4, Nanog, and Sox-2 (ONS) expression. Similar to embryonic stem cells (ESCs), high ONS expression eventually became the measure of pluripotency in any cell. The threshold expression of ONS genes that underscores pluripotency, stemness, and differentiation potential is still unclear. Therefore, we raised a question as to whether pluripotency and stemness is a function of basal ONS gene expression. To prove this, we carried out a comparative study between basal ONS expressing NIH3T3 cells with pluripotent mouse bone marrow mesenchymal stem cells (mBMSC) and mouse ESC. Our studies on cellular, molecular, and immunological biomarkers between NIH3T3 and mBMSC demonstrated stemness property of undifferentiated NIH3T3 cells that was similar to mBMSC and somewhat close to ESC as well. In vivo teratoma formation with all three germ layer derivatives strengthen the fact that these cells in spite of basal ONS gene expression can differentiate into cells of multiple lineages without any genetic modification. Conclusively, our novel findings suggested that the phenomenon of pluripotency which imparts ability for multilineage cell differentiation is not necessarily a function of high ONS gene expression.

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Section: In Vivo Teratoma Formation Reconfirms Pluripotency In Nih3t3supporting

confidence: 91%

Basal Expression of Pluripotency-Associated Genes Can Contribute to Stemness Property and Differentiation Potential

Dadheech

Srivastava

Belani

et al. 2013

Stem Cells and Development

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“…The epidermis is a dynamic epithelium that is constantly renewed throughout life. A subpopulation of cell, epidermal progenitor cells are responsibility for the epidermis self-renew, which is located in the upper hair follicle (bulge) and patches of basal keratinocytes [28]–[30]. There are now a number of reports indicating that epidermal progenitor cells have differentiation potential outside the organ in which they reside [31], [32].…”

Section: Discussionmentioning

confidence: 99%

“…We isolated monkey epidermal progenitor cells with type IV collagen adhesion, in the most effective procedure used to isolate epidermal progenitor cells [18], [30], [35], [36]. Type IV collagen, the β 1 Integrin ligand, is responsible for the attachment of the basal layer of the epidermis to its underlying substratum, the basement membrane.…”

Section: Discussionmentioning

confidence: 99%

Conjunctival Reconstruction with Progenitor Cell-Derived Autologous Epidermal Sheets in Rhesus Monkey

Zhang

Huang

et al. 2011

PLoS ONE

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Severe ocular surface diseases are some of the most challenging problems that the clinician faces today. Conventional management is generally unsatisfactory, and the long-term ocular consequences of these conditions are devastating. It is significantly important to find a substitute for conjunctival epithelial cells. This study was to explore the possibility of progenitor cell-derived epidermal sheets on denuded amniotic membrane to reconstruct ocular surface of conjunctiva damaged monkeys. We isolated epidermal progenitor cells of rhesus monkeys by type IV collagen adhesion, and then expanded progenitor cell-derived epidermal sheets on denuded amniotic membrane ex vivo. At 3 weeks after the conjunctiva injury, the damaged ocular surface of four monkeys was surgically reconstructed by transplanting the autologous cultivated epidermal progenitor cells. At 2 weeks after surgery, transplants were removed and examined with Hematoxylin-eosin staining, Periodic acid Schiff staining, immunofluorescent staining, scanning and transmission electron microscopy. Histological examination of transplanted sheets revealed that the cell sheets were healthy alive, adhered well to the denuded amniotic membrane, and had several layers of epithelial cells. Electron microscopy showed that the epithelial cells were very similar in appearance to those of normal conjunctival epithelium, even without goblet cell detected. Epithelial cells of transplants had numerous desmosomal junctions and were attached to the amniotic membrane with hemidesmosomes. Immunohistochemistry confirmed the presence of the conjunctival specific markers, mucin 4 and keratin 4, in the transplanted epidermal progenitor cells. In conclusion, our present study successfully reconstructed conjunctiva with autologous transplantation of progenitor cell-derived epidermal sheets on denuded AM in conjunctival damaged monkeys, which is the first step toward assessing the use of autologous transplantation of progenitor cells of nonocular surface origin. Epidermal progenitor cells could be provided as a new substitute for conjunctival epithelial cells to overcome the problems of autologous conjunctiva shortage.

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“…The rapidly adhering cells were collected and cultured in K-SFM medium at 37 °C in 5% CO 2 . When the culture reached 70–80% confluence, the cells were digested and passaged at a ratio of 1:2 [16]. Meanwhile, the cells were identified to be ESCs with integrin-α6 bri (3750S; CST, BOS, USA) and CD71 dim (553264; BD) by immunofluorescence staining [17].…”

Section: Methodsmentioning

confidence: 99%

Basic fibroblast growth factor reduces scar by inhibiting the differentiation of epidermal stem cells to myofibroblasts via the Notch1/Jagged1 pathway

Wang

Shu

et al. 2017

Stem Cell Res Ther

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BackgroundBasic fibroblast growth factor (bFGF) plays an important role in promoting wound healing and reducing scar, but the possible molecular mechanisms are still unclear. Our previous studies have found that activating the Notch1/Jagged1 pathway can inhibit the differentiation of epidermal stem cells (ESCs) to myofibroblasts (MFB). Herein, we document that bFGF reduces scar by inhibiting the differentiation of ESCs to MFB via activating the Notch1/Jagged1 pathway.MethodsIn in-vitro study, ESCs were isolated from 10 neonatal SD rats (1–3 days old), cultured in keratinocyte serum-free medium, and divided into six groups: bFGF group, bFGF + SU5402 group, bFGF + DAPT group, siJagged1 group, bFGF + siJagged1 group, and control group. Jagged1 of the ESCs in the siJagged1 group and bFGF + siJagged1 group was knocked down by small-interfering RNA transfection. Expression of ESC markers (CK15/CK10), MFB markers (α-SMA, Collagen I, Collagen III), and Notch1/Jagged1 components (Jagged1, Notch1, Hes1) was detected by FCM, qRT-PCR, and western blot analysis to study the relationships of bFGF, ESCs, and Notch1/Jagged1 pathway. In in-vivo study, the wound healing time and scar hyperplasia were observed on rabbit ear scar models. The quality of wound healing was estimated by hematoxylin and eosin staining and Masson staining. Expression of ESC markers, MFB markers and Notch1/Jagged1 components was elucidated by immunohistochemistry, immunofluorescence, and western blot analysis.ResultsThe in-vitro study showed that bFGF could significantly upregulate the expression of ESC markers and Notch1/Jagged1 components, while downregulating the expression of MFB markers at the same time. However, these effects could be obviously decreased when we knocked down Jagged1 or added DAPT. Similarly, in in-vivo study, bFGF also exhibited its functions in inhibiting the differentiation of rabbit ESCs to MFB by activating the Notch1/Jagged1 pathway, which improved the wound healing quality and alleviated scar significantly.ConclusionThese results provide evidence that bFGF can reduce scar by inhibiting the differentiation of ESCs to MFB via the Notch1/Jagged1 pathway, and present a new promising potential direction for the treatment of scar.

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Isolation, culture and identification of epidermal stem cells from newborn mouse skin

Cited by 25 publications

References 45 publications

Basal Expression of Pluripotency-Associated Genes Can Contribute to Stemness Property and Differentiation Potential

Basal Expression of Pluripotency-Associated Genes Can Contribute to Stemness Property and Differentiation Potential

Conjunctival Reconstruction with Progenitor Cell-Derived Autologous Epidermal Sheets in Rhesus Monkey

Basic fibroblast growth factor reduces scar by inhibiting the differentiation of epidermal stem cells to myofibroblasts via the Notch1/Jagged1 pathway

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