Simultaneous Isolation of Three Different Stem Cell Populations from Murine Skin

Forni, Maria Fernanda; Lobba, Aline Ramos Maia; Ferreira, Aryel H.; Sogayar, Mari Cleide

doi:10.1371/journal.pone.0140143

Cited by 15 publications

(11 citation statements)

References 31 publications

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“…Cells were obtained as described in ref. . Briefly, subcutaneous fat was removed from the skin with a scalpel, and the whole skin was placed dermis‐down in trypsin (Gibco) at 37 °C for 30 minutes.…”

Section: Methodsmentioning

confidence: 99%

“…Cells were fixed in 4% w/v paraformaldehyde and subjected to immunofluorescence microscopy as previously described . The antibodies and their dilutions are listed in Supporting Information Table 2.…”

Section: Methodsmentioning

confidence: 99%

“…Mesenchymal stem cells (MSCs) are found in virtually all post‐natal organs and tissues and display the ability to differentiate in vitro and in vivo into bone, cartilage, and adipose tissues, among others . Mouse skin mesenchymal stem cells (msMSCs) are CD105 + CD90 + CD73 + CD29 + CD34 − mesodermal precursors located in the dermis that, after induction, display classic differentiation potential in vitro . Recent studies suggest that extensive metabolic reconfiguration occurs during differentiation.…”

Section: Introductionmentioning

confidence: 99%

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Murine Mesenchymal Stem Cell Commitment to Differentiation Is Regulated by Mitochondrial Dynamics

et al. 2015

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Mouse skin mesenchymal stem cells (msMSCs) are dermis CD105+CD90+CD73+CD29+CD34− mesodermal precursors which, after in vitro induction, undergo chondro, adipo and osteogenesis. Extensive metabolic reconfiguration has been found to occur during differentiation, and the bioenergetic status of a cell is known to be dependent on the quality and abundance of the mitochondrial population, which may be regulated by fusion and fission. However, little is known regarding the impact of mitochondrial dynamics on the differentiation process. We addressed this knowledge gap by isolating MSCs from Swiss female mice, inducing these cells to differentiate into osteo, chondro and adipocytes and measuring changes in mass, morphology, dynamics and bioenergetics. Mitochondrial biogenesis was increased in adipogenesis, as evaluated through confocal microscopy, citrate synthase activity and mtDNA content. The early steps of adipo and osteogenesis involved mitochondrial elongation, as well as increased expression of mitochondrial fusion proteins Mfn1 and 2. Chondrogenesis involved a fragmented mitochondrial phenotype, increased expression of fission proteins Drp1, Fis1 and 2 and enhanced mitophagy. These events were accompanied by profound bioenergetic alterations during the commitment period. Moreover, knockdown of Mfn2 in adipo and osteogenesis and the overexpression of a dominant negative form of Drp1 during chondrogenesis resulted in a loss of differentiation ability. Overall, we find that mitochondrial morphology and its regulating processes of fission/fusion are modulated early on during commitment, leading to alterations in the bioenergetic profile that are important for differentiation. We thus propose a central role for mitochondrial dynamics in the maintenance/commitment of mesenchymal stem cells.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Murine Mesenchymal Stem Cell Commitment to Differentiation Is Regulated by Mitochondrial Dynamics

et al. 2015

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“…The adherent cell colonies achieved from culture condition B displayed the most important markers and characteristics that a primary culture should possess to be classified as DMSCs [ 24 ]. Forni et al [ 25 ] reported simultaneously isolating and characterizing three stem cell populations from the dermis and epidermis of murine skin, namely epidermal stem cells, SKPs, and DMSCs. They believed DMSCs and SKPs could be isolated simultaneously from one skin sample and showed no overt difference with DMSCs and SKPs isolated in a routine way.…”

Section: Discussionmentioning

confidence: 99%

Facilitation of mouse skin-derived precursor growth and yield by optimizing plating density

Xiong

Dai

et al. 2021

Open Life Sciences

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Multiple methodologies have been reported to facilitate skin-derived precursor (SKP) growth, but the impact of plating density on SKP growth has not been studied. To determine the optimal plating density, we used six plating densities and two types of flasks for mouse SKP (mSKP) culture. On the 14th day, the number, diameter, and viability of mSKP spheres were compared by morphological assessment and cell counting kit 8, and we found the optimal plating density was 2.5 × 105–5 × 105 cells/mL. In addition, we investigated the correlation between the SKP spheres and the adherent cell colonies in the serum-free culture system. We treated the adherent cell colonies with two culture conditions and characterized the cells generated from two conditions by immunocytochemistry and induced differentiation, respectively. The results elucidated that the adherent cell colonies differentiated into either mSKPs or dermal mesenchymal stem cells under appropriate culture conditions. In conclusion, mSKP spheres differentiated from the adherent cell colonies. The optimal plating density significantly promoted and advanced the proliferation of adherent cell colonies, which optimized mSKP growth and yield. The adherent cell colonies possessed the capacity of differentiating into different types of cells under appropriate culture conditions.

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“…As there is a positive correlation between the IF stem cell proliferation and adherence, the proliferative IF stem cells can be distinguished from the low-adhered TA cells with the higher β1 integrins expression [20]. During terminal differentiation, the IF stem cells express involucrin, a differentiation marker, and filaggrin, an intermediate filament (IF)-associated protein [21,22].…”

Section: If Stem Cell Markersmentioning

confidence: 99%

Keratinocyte Stem Cells: Role in Aging

Ansary¹,

Kamiya²,

Ohtsuki³

2022

Keratinocyte Biology - Structure and Function in the Epidermis

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Stem cells located in the skin are responsible for continual regeneration, wound healing, and differentiation of different cell lineages of the skin. The three main locations of skin stem cells are the epidermis, dermis, and hair follicles. The keratinocyte stem cells are located in the epidermal basal layer (the interfollicular stem cells), hair follicle bulge region (the hair follicle stem cells), and sebaceous glands (the sebaceous gland stem cells) and are responsible for the epidermal proliferation, differentiation, and apoptosis. The interfollicular (IF) stem cells are responsible for epidermis regeneration by proliferating basal cells that attach to the underlying basement membrane and with time they exit from the cell cycle, start terminal differentiation, and move upward to form the spinous, the granular, and the stratum corneum layers. The hair follicle (HF) stem cells are responsible for hair regeneration and these stem cells undergo a cycle consists three stages; growth cycles (anagen), degeneration (catagen), and relative resting phase (telogen). The sebaceous gland (SG) stem cells located in between the hair follicle bulge and the gland and are responsible for producing the entire sebaceous gland which secretes oils to moisture our skin. The role of epidermal stem cells is extremely crucial because they produce enormous numbers of keratinocytes over a lifetime to maintain epidermal homeostasis. However, the age-associated changes in the skin; for example; alopecia, reduced hair density, gray or thin hair, reduced wound healing capacity are related to skin stem cells’ decline functionality with age.

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Simultaneous Isolation of Three Different Stem Cell Populations from Murine Skin

Cited by 15 publications

References 31 publications

Murine Mesenchymal Stem Cell Commitment to Differentiation Is Regulated by Mitochondrial Dynamics

Murine Mesenchymal Stem Cell Commitment to Differentiation Is Regulated by Mitochondrial Dynamics

Facilitation of mouse skin-derived precursor growth and yield by optimizing plating density

Keratinocyte Stem Cells: Role in Aging

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