Adult skin-derived precursor Schwann cells exhibit superior myelination and regeneration supportive properties compared to chronically denervated nerve-derived Schwann cells

Kumar, Ranjan; Sinha, Sarthak; Hagner, Andrew; Stykel, Morgan G.; Raharjo, Eko; Singh, Karun K.; Midha, Rajiv; Biernaskie, Jeff

doi:10.1016/j.expneurol.2016.02.006

Cited by 27 publications

(30 citation statements)

References 91 publications

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“…Using NES as a reliable marker of SCs in vitro (Kumar et al, 2016), we performed coexpression analysis of several additional proteins in nerve and skin-derived SC cultures as well as patient-matched dermal fibroblast cultures (Figs. 3, 4).…”

Section: Resultsmentioning

confidence: 99%

“…To assess the genetic profile of adult human SCs and fibroblasts, cells were allowed to grow in SC media until 80% confluent, or in differentiation/promyelination media (50 μg/ml ascorbic acid, 0.5 μM forskolin, 5 ng/ml neuregulins, and 5 ng/ml NGF) for 14 d on axons grown as previously described (Kumar et al, 2016). They were then washed with sterile PBS and 500,000 cells were pelleted, and snap frozen.…”

Section: Methodsmentioning

confidence: 99%

“…All rodents were kept in a 12/12 h light/dark cycle, temperature-controlled environment with unlimited food and water. All procedures were done as previously described (McKenzie et al, 2006; Kumar et al, 2016). Briefly, sciatic nerves were exposed, and a mid-thigh level crush was induced using number 5 surgical forceps for 60 s while mice were under deep anesthetic (2% isoflurane).…”

Section: Methodsmentioning

confidence: 99%

“…An alternative approach might be to source SCs from a more accessible source, such as the skin (Wong et al, 2006; Biernaskie et al, 2006). Bona fide SCs can be differentiated in vitro from isolated, self-renewing skin-derived precursors (SKPs) and following transplantation have been shown to improve anatomic measures such as axon growth, myelin clearance, modulation of endogenous glial cell behavior, and remyelination, ultimately supporting improved behavioral outcomes following spinal cord and nerve injury (McKenzie et al, 2006; Biernaskie et al, 2007; Walsh et al, 2009; Walsh et al, 2010; Khuong et al, 2014; Sparling et al, 2015; Kumar et al, 2016; Stratton et al, 2016). Although the majority of these studies have been done only using rodent cells, recent work shows myelination by human skin-derived SCs (hSk-SCs) is possible from the foreskin of neonatal children (Krause et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

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Purification and Characterization of Schwann Cells from Adult Human Skin and Nerve

Stratton

Kumar

Sinha

et al. 2017

eNeuro

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Despite its modest capacity for regeneration, peripheral nervous system injury often results in significant long-term disability. Supplementing peripheral nervous system injury with autologous Schwann cells (SCs) may serve to rejuvenate the postinjury environment to enhance regeneration and ultimately improve functional outcomes. However, human nerve-derived SC (hN-SC) collection procedures require invasive surgical resection. Here, we describe the characterization of SCs from adult human skin (hSk-SCs) of four male donors ranging between 27 and 46 years old. Within five weeks of isolating and culturing adherent mixed skin cells, we were able to obtain 3–5 million purified SCs. We found that hSk-SCs appeared transcriptionally indistinguishable from hN-SCs with both populations exhibiting expression of SC genes including: SOX10, SOX9, AP2A1, CDH19, EGR1, ETV5, PAX3, SOX2, CX32, DHH, NECL4, NFATC4, POU3F1, S100B, and YY1. Phenotypic analysis of hSk-SCs and hN-SCs cultures revealed highly enriched populations of SCs indicated by the high percentage of NES+ve, SOX10+ve, s100+ve and p75+ve cells, as well as the expression of a battery of other SC-associated proteins (PAX3, CDH19, ETV5, SOX2, POU3F1, S100B, EGR2, and YY1). We further show that both hSk-SCs and hN-SCs are capable of promoting axonal growth to similar degrees and that a subset of both associate with regenerating axons and form myelin following transplantation into the injured mouse sciatic nerve. Interestingly, although the majority of both hSk-SCs and hN-SCs maintained SOX10 immunoreactivity following transplant, only a subset of each activated the promyelinating factor, POU3F1, and were able to myelinate. Taken together, we demonstrate that adult hSk-SCs are genetically and phenotypically indistinguishable to hN-SCs.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Purification and Characterization of Schwann Cells from Adult Human Skin and Nerve

Stratton

Kumar

Sinha

et al. 2017

eNeuro

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“…Over the past decade, there has been extensive development of tissue engineering techniques that provide a new way for enhancing peripheral nerve regeneration and functional recovery33. Peripheral nerve repair with tissue-engineered technology requires a large quantity of seed cells.…”

Section: Discussionmentioning

confidence: 99%

Human eyelid adipose tissue-derived Schwann cells promote regeneration of a transected sciatic nerve

Wang

Cao

Wang

et al. 2017

Sci Rep

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Schwann cells (SCs) can promote the regeneration of injured peripheral nerves while the clinical application is limited by donor site complications and the inability to generate an ample amount of cells. In this study, we have isolated human eyelid adipose-derived Schwann cells (hE-SCs) from human eyelid adipose tissue and identified the cell phenotype and function. Using immunofluorescence and H & E staining, we detected subtle nerve fibers and SCs in human eyelid adipose tissue. Immunofluorescence staining indicated that hE-SCs expressed glial markers, such as S100, p75NTR GFAP, Sox10 and Krox20. To explore whether hE-SCs promote the regeneration of injured peripheral nerves in vivo, a Balb/c-nu mice model was used in the study, and mice were randomly assigned to five groups: Matrigel; hE-SCs/P0; hE-SCs/P2; hE-FLCs/P2; and Autograft. After 12 weeks, functional and histological assessments of the regenerated nerves showed that sciatic nerve defect was more effectively repaired in the hE-SCs/P2 group which achieved 66.1 ± 6.5% purity, than the other three groups and recovered to similar level to the Autograft group. These results indicated that hE-SCs can promote the regeneration of injured peripheral nerve and the abundant, easily accessible supply of adipose tissue might be a promising source of SCs for peripheral nerve repair.

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Control of dissolved oxygen significantly increases the yield of skin‐derived Schwann cells during expansion in stirred suspension bioreactors

Walsh

Biernaskie

Midha

et al. 2021

Engineering Reports

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Peripheral nerve and spinal cord injuries affect millions of people worldwide often leading to lifelong disability and pain. Cell therapy using autologous Schwann cells are currently being investigated for the treatment of these injuries and are showing great potential. In particular, skin-derived Schwann cells offer a non-invasive option for highly therapeutic cells that have been shown to regenerate tissue and restore function. However, it is estimated that for an effective therapy, hundreds of millions of Schwann cells are needed, and current static and spinner flask methods cannot generate these clinical numbers. Therefore, this study utilizes control systems facilitated by stirred suspension bioreactors to monitor and maintain the pH and dissolved oxygen at levels that promote cell proliferation and survival. By adapting the current culture practices to scalable bioreactor platforms, we were able to control the dissolved oxygen creating new expansion protocols that enabled the bioreactor process to grow to clinical scale, producing 667 × 10 6 cells in a single 500 mL bioreactor in 7 days. Since individual injury is different in terms of complexity and severity, and therefore cell needs, by changing the available surface area in the bioreactor, this developed process is highly adjustable to meet the cell quantity needs for each individual treatment. Following the bioprocess, cells that were harvested from the microcarriers continue to express typical Schwann cell markers suggesting that they retain phenotypic stability. By increasing the production potential of skin-derived Schwann cells, this therapy is one step closer to the clinic with the hope for ultimately restoring function and improving quality of life to patients with neural injuries.

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Adult skin-derived precursor Schwann cells exhibit superior myelination and regeneration supportive properties compared to chronically denervated nerve-derived Schwann cells

Cited by 27 publications

References 91 publications

Purification and Characterization of Schwann Cells from Adult Human Skin and Nerve

Purification and Characterization of Schwann Cells from Adult Human Skin and Nerve

Human eyelid adipose tissue-derived Schwann cells promote regeneration of a transected sciatic nerve

Control of dissolved oxygen significantly increases the yield of skin‐derived Schwann cells during expansion in stirred suspension bioreactors

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