Stem/Progenitor Cells in Non-Lactating Versus Lactating Equine Mammary Gland

Spaas, Jan H.; Chiers, Koen; Bussche, Leen; Burvenich, Christian; Walle, Gerlinde R. Van de

doi:10.1089/scd.2012.0042

Cited by 17 publications

(20 citation statements)

References 66 publications

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“…Finding suitable antibodies is, therefore, a challenge in equine studies. Adipogenic differentiation is usually described after 21 days for MSCs [5]; however, recently, adipogenesis of mammary stem/progenitor cells has been reported as soon as 5 days [66] and even 3 days in the present study. If MSCs would be stained after 3 days of adipogenic differentiation under the same circumstances, adipocytes might also be detected.…”

Section: Discussioncontrasting

confidence: 49%

Equine Epidermis: A Source of Epithelial-Like Stem/Progenitor Cells with In Vitro and In Vivo Regenerative Capacities

Broeckx¹,

Maes²,

Martinello

et al. 2014

Stem Cells and Development

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Besides the presence of somatic stem cells in hair follicles and dermis, the epidermis also contains a subpopulation of stem cells, reflecting its high regenerative capacity. However, only limited information concerning epidermis-derived epithelial-like stem/progenitor cells (EpSCs) is available to date. Nonetheless, this stem cell type could prove itself useful in skin reconstitution after injury. After harvesting from equine epidermis, the purified cells were characterized as EpSCs by means of positive expression for CD29, CD44, CD49f, CD90, Casein Kinase 2b, p63, and Ki67, low expression for cytokeratin (CK)14 and negative expression for CD105, CK18, Wide CK, and Pan CK. Furthermore, their self-renewal capacity was assessed in adhesion as well as in suspension. Moreover, the isolated cells were differentiated toward keratinocytes and adipocytes. To assess the regenerative capacities of EpSCs, six full-thickness skin wounds were made: three were treated with EpSCs and platelet-rich-plasma (EpSC/PRP-treated), while the remaining three were administered carrier fluid alone (PRP-treated). The dermis of EpSC/PRP-treated wounds was significantly thinner and exhibited more restricted granulation tissue than did the PRP-treated wounds. The EpSC/PRP-treated wounds also exhibited increases in EpSCs, vascularization, elastin content, and follicle-like structures. In addition, combining EpSCs with a PRP treatment enhanced tissue repair after clinical application.

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Section: Discussioncontrasting

confidence: 49%

Equine Epidermis: A Source of Epithelial-Like Stem/Progenitor Cells with In Vitro and In Vivo Regenerative Capacities

Broeckx¹,

Maes²,

Martinello

et al. 2014

Stem Cells and Development

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“…Moreover, the authors assessed their clonogenic expansion capacities using CFU assays and studied equine MaSC in more detail during lactation. Hereby, it was observed that equine lactating mammary glands contained significantly more mammosphere-initiating cells than the inactive, nonlactating gland (a reflection of MaSC self-renewal) and moreover, that these spheres were significantly larger in size upon initial cultivation (a reflection of progenitor cell proliferation) [45].…”

Section: Equine Mascmentioning

confidence: 93%

“…The sphere formation efficiency of serially cultivated cells can be used as a self-renewal test. To this end, a limited number of cells is seeded at a clonal density and cultivated for 7 days, after which the number of colony-forming units (CFU) is evaluated [45]. For the differentiation test, cells are cultured under differentiation conditions and immunostained with lineage-specific markers.…”

Section: Mammospheres Self-renewal and Differentiation Testsmentioning

confidence: 99%

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Mammary Stem Cell Research in Veterinary Science: An Update

Borena

Bussche

Burvenich

et al. 2013

Stem Cells and Development

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The mammary gland is an organ with a remarkable regenerative capacity that can undergo multiple cycles of proliferation, lactation, and involution. Growing evidence suggests that these changes are driven by the coordinated division and differentiation of mammary stem cell populations (MaSC). Whereas information regarding MaSC and their role in comparative mammary gland physiology is readily available in human and mice, such information remains scarce in most veterinary mammal species such as cows, horses, sheep, goats, pigs, and dogs. We believe that a better knowledge on the MaSC in these species will not only help to gain more insights into mammary gland (patho) physiology in veterinary medicine, but will also be of value for human medicine. Therefore, this review summarizes the current knowledge on stem cell isolation and characterization in different mammals of veterinary importance.

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“…In the mammary gland, another epidermal organ, so-called mammospheres are also intensively studied for stem/progenitor cell purification purposes [26,27]. Unfortunately, a major disadvantage of this 3-D technique is contamination with other adult cell types surviving between the sphere-forming stem cells [26].…”

Section: Introductionmentioning

confidence: 99%

Sphere-Forming Capacity as an Enrichment Strategy for Epithelial-Like Stem Cells from Equine Skin

Borena

Meyer

Chiers

et al. 2014

Cell Physiol Biochem

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Background: Mammal skin plays a pivotal role in several life preserving processes and extensive damage may therefore be life threatening. Physiological skin regeneration is achieved through ongoing somatic stem cell differentiation within the epidermis and the hair follicle. However, in severe pathological cases, such as burn wounds, chronic wounds, and ulcers, the endogenous repair mechanisms might be insufficient. For this reason, exogenous purification and multiplication of epithelial-like stem/progenitor cells (EpSCs) might be useful in the treatment of these skin diseases. However, only few reports are available on the isolation, purification and characterization of EpSCs using suspension cultures. Methods: In the present study, skin was harvested from 6 mares and EpSCs were isolated and purified. In addition to their characterization based on phenotypic and functional properties, sphere formation was assessed upon isolation, i.e. at passage 0 (P₀), and at early (P₄) and late (P₁₀) passages using different culture conditions. Results: On average 0.53 ± 0.28% of these primary skin-derived cells showed the capacity to form spheres and hence possessed stem cell properties. Moreover, significantly more spheres were observed in EpSC medium versus differentiation medium, corroborating the EpSCs' privileged ability to survive in suspension. Furthermore, the number of cells per sphere significantly increased over time as well as with subsequent passaging. Upon immunophenotyping, the presumed EpSCs were found to co-express cytokeratin (CK) 14, Casein kinase 2 beta and Major Histocompatibility Complex (MHC) I and expressed no pan CK and wide CK. Only a few cells expressed MHC II. Their differentiation towards keratinocytes (at P₄ and P₁₀) was confirmed based on co-expression of CK 14, Casein kinase 2 beta, pan CK and wide CK. In one of six isolates, a non-EpSC cell type was noticed in adherent culture. Although morphological features and immunohistochemistry (IHC) confirmed a keratinocyte phenotype, this culture could be purified by seeding the cells in suspension at ultralow clonal densities (1 and 10 cells/cm²), yet with a significantly lower sphere forming efficiency in comparison to pure EpSCs (P = 0.0012). Conclusion: The present study demonstrated sphere formation as a valuable tool to purify EpSCs upon their isolation and assessed its effectiveness at different clonal seeding densities for eliminating a cellular contamination.

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Stem/Progenitor Cells in Non-Lactating Versus Lactating Equine Mammary Gland

Cited by 17 publications

References 66 publications

Equine Epidermis: A Source of Epithelial-Like Stem/Progenitor Cells with In Vitro and In Vivo Regenerative Capacities

Equine Epidermis: A Source of Epithelial-Like Stem/Progenitor Cells with In Vitro and In Vivo Regenerative Capacities

Mammary Stem Cell Research in Veterinary Science: An Update

Sphere-Forming Capacity as an Enrichment Strategy for Epithelial-Like Stem Cells from Equine Skin

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