Isolation and Characterization of Satellite Cells from Rat Head Branchiomeric Muscles

Monroy, Paola L. Carvajal; Yablonka–Reuveni, Zipora; Grefte, Sander; Kuijpers-Jagtman, Anne Marie; Wagener, Frank A. D. T. G.; Hoff, Johannes W. Von den

doi:10.3791/52802

Cited by 7 publications

(7 citation statements)

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“…However, velopharyngeal dysfunction persists in 7% to 30% of treated patients, and deficiencies in speech are common (Marrinan et al 1998; Inman et al 2005; Mossey et al 2009). Several factors contribute to the poor outcomes associated with surgery, but fibrosis seems to be an important factor (Carvajal Monroy et al 2012; Carvajal Monroy, Grefte, et al 2015).…”

Section: Introductionmentioning

confidence: 99%

Neonatal Satellite Cells Form Small Myotubes In Vitro

et al. 2016

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Although palatal muscle reconstruction in patients with cleft palate takes place during early childhood, normal speech development is often not achieved. We hypothesized that the intrinsic properties of head satellite cells (SCs) and the young age of these patients contribute to the poor muscle regeneration after surgery. First, we studied the fiber type distribution and the expression of SC markers in ex vivo muscle tissue from head (branchiomeric) and limb (somite-derived) muscles from neonatal (2-wk-old) and young (9-wk-old) rats. Next, we cultured SCs isolated from these muscles for 5, 7, and 9 d, and investigated the in vitro expression of SC markers, as well as changes in proliferation, early differentiation, and fusion index (myotube formation) in these cells. In our ex vivo samples, we found that virtually all myofibers in both the masseter (Mass) and the levator veli palatini (LVP) muscles contained fast myosin heavy chain (MyHC), and a small percentage of digastric (Dig) and extensor digitorum longus myofibers also contained slow MyHC. This was independent of age. More SCs were found in muscles from neonatal rats as compared with young rats [17.6 (3.8%) v. 2.3 (1.6%); P < 0.0001]. In vitro, young branchiomeric head muscle (BrHM) SCs proliferated longer and differentiated later than limb muscle SCs. No differences were found between SC cultures from the different BrHMs. SC cultures from neonatal muscles showed a much higher proliferation index than those from young animals at 5 d (0.8 v. 0.2; P < 0.001). In contrast, the fusion index in neonate SCs was about twice as low as that in SCs from young muscles at 9 d [27.6 (1.4) v. 62.8 (10.2), P < 0.0001]. In conclusion, SCs from BrHM differ from limb muscles especially in their delayed differentiation. SCs from neonatal muscles form myotubes less efficiently than those from young muscles. These age-dependent differences in stem cell properties urge careful consideration for future clinical applications in patients with cleft palate.

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Section: Introductionmentioning

confidence: 99%

Neonatal Satellite Cells Form Small Myotubes In Vitro

et al. 2016

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“…Next, the masseter muscles from both sides of the head were dissected and stored in phosphate‐buffered saline (PBS; Gibco) supplemented with penicillin/streptomycin 2% (Gibco). Satellite cells (SCs) were isolated as described in a previous publication [ 44 ]. In short, the muscles were minced and digested in Pronase 0.1% (Calbiochem, Sigma‐Aldrich) for 1 h at 37°C, pipetted up and down multiple times and centrifuged.…”

Section: Methodsmentioning

confidence: 99%

“…The isolated cells were pooled, expanded for 3 d in culture medium and frozen in 1 mL vials for later use. Each masseter yields approximately 1 × 10 6 cells [ 44 ]. After a 1‐day culture on matrigel, this cell population contained 80.0% [SD 13.8%] Pax7‐positive cells.…”

Section: Methodsmentioning

confidence: 99%

“…The culture medium was Dulbecco's modified Eagle's medium with 4,500 mg/L glucose, 4 mL L‐glutamine, and 110 mg/mL sodium pyruvate supplemented with 20% fetal bovine serum (v/v), 10% horse serum (v/v), 1% penicillin‐streptomycin, and 1% chicken extract embryo filtered through a 0.2 μm filter. This culture medium promotes both the proliferation as well as differentiation of myoblasts [ 44 , 45 ]. The medium was changed every day.…”

Section: Methodsmentioning

confidence: 99%

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Effect of niche components on masseter satellite cell differentiation on fibrin coatings

Lijten

Salazar

Erp

et al. 2022

European J Oral Sciences

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In skeletal muscles, niche factors stimulate satellite cells to activate and induce muscle regeneration after injury. In vitro, matrigel is widely used for myoblast differentiation, however, is unsuitable for clinical applications. Therefore, this study aimed to analyze attachment and differentiation of satellite cells into myotubes on fibrin coatings with selected niche components. The attachment of satellite cells to fibrin alone and fibrin with niche components (laminin, collagen-IV, laminin-entactin complex [LEC]) were compared to matrigel. Only on matrigel and fibrin with LEC, Pax7-positive cells attached well. Then, LEC was selected to analyze proliferation, differentiation, and fusion indices. The proliferation index at day 1 on fibrin-LEC (22.5%, SD 9.1%) was similar to that on matrigel (30.8% [SD 11.1%]). The differentiation index on fibrin-LEC (28.7% [SD 6.1%] at day 5 and 32.8% [SD 6.7%] at day 7) was similar to that on matrigel (40.1% [5.1%] at day 5 and 27.1% [SD 4.3%] at day 7). On fibrin-LEC, the fusion index at day 9 (26.9% [SD 11.5%]) was similar to that on matrigel (25.5% [SD 4.7%]). Our results showed that the addition of LEC enhances the formation of myotubes on fibrin. Fibrin with LEC might be suitable to enhance muscle regeneration after surgery such as cleft palate repair and other muscle defects.

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“…Muscle repair following injury is facilitated by SCs. Satellite cells are muscle stem cells originating from the CPM that are responsible for postnatal muscle growth, maintenance, and repair . They express the paired box transcription factor 7 (Pax7) and are located between the sarcolemma and basal lamina surrounding a single muscle fiber.…”

Section: Stem Cells Repair Wounds and Facilitate Skin And Muscle Regementioning

confidence: 99%

Tissue engineering strategies combining molecular targets against inflammation and fibrosis, and umbilical cord blood stem cells to improve hampered muscle and skin regeneration following cleft repair

Schreurs

Suttorp

Mutsaers

et al. 2019

Medicinal Research Reviews

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Cleft lip with or without cleft palate is a congenital deformity that occurs in about 1 of 700 newborns, affecting the dentition, bone, skin, muscles and mucosa in the orofacial region. A cleft can give rise to problems with maxillofacial growth, dental development, speech, and eating, and can also cause hearing impairment. Surgical repair of the lip may lead to impaired regeneration of muscle and skin, fibrosis, and scar formation. This may result in hampered facial growth and dental development affecting oral function and lip and nose esthetics. Therefore, secondary surgery to correct the scar is often indicated. We will discuss the molecular and cellular pathways involved in facial and lip myogenesis, muscle anatomy in the normal and cleft lip, and complications following surgery. The aim of this review is to outline a novel molecular and cellular strategy to improve musculature and skin regeneration and to reduce scar formation following cleft repair. Orofacial clefting can be diagnosed in the fetus through prenatal ultrasound screening and allows planning for the harvesting of umbilical cord blood stem cells upon birth. Tissue engineering techniques using these cord blood stem cells and molecular targeting of inflammation and fibrosis during surgery may promote tissue regeneration. We expect that this novel strategy improves both muscle and skin regeneration, resulting in better function and esthetics after cleft repair.

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Isolation and Characterization of Satellite Cells from Rat Head Branchiomeric Muscles

Cited by 7 publications

References 50 publications

Neonatal Satellite Cells Form Small Myotubes In Vitro

Neonatal Satellite Cells Form Small Myotubes In Vitro

Effect of niche components on masseter satellite cell differentiation on fibrin coatings

Tissue engineering strategies combining molecular targets against inflammation and fibrosis, and umbilical cord blood stem cells to improve hampered muscle and skin regeneration following cleft repair

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