Biodegradable ECM-coated PCL microcarriers support scalable human early MSC expansion and in vivo bone formation

Shekaran, Asha; Lam, Alan Tin-Lun; Sim, Eileen; Lee, Jialing; Li, Jian; Wen, Jessica Toh Pei; Chan, Jerry Kok Yen; Choolani, Mahesh; Reuveny, Shaul; Birch, William R.; Oh, Steve

doi:10.1016/j.jcyt.2016.06.016

Cited by 60 publications

(49 citation statements)

References 23 publications

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“…In addition, we have also shown that biodegradable heMSC-PCL microcarrier constructs can be generated and transplanted in vivo for bone regeneration [30]. This advantage of utilizing microcarriers to improve the differentiation potential of MSC is further supported by work from other groups [31–33].…”

Section: Introductionmentioning

confidence: 90%

Critical attributes of human early mesenchymal stromal cell-laden microcarrier constructs for improved chondrogenic differentiation

et al. 2017

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BackgroundMicrocarrier cultures which are useful for producing large cell numbers can act as scaffolds to create stem cell-laden microcarrier constructs for cartilage tissue engineering. However, the critical attributes required to achieve efficient chondrogenic differentiation for such constructs are unknown. Therefore, this study aims to elucidate these parameters and determine whether cell attachment to microcarriers throughout differentiation improves chondrogenic outcomes across multiple microcarrier types.MethodsA screen was performed to evaluate whether 1) cell confluency, 2) cell numbers, 3) cell density, 4) centrifugation, or 5) agitation are crucial in driving effective chondrogenic differentiation of human early mesenchymal stromal cell (heMSC)-laden Cytodex 1 microcarrier (heMSC-Cytodex 1) constructs.ResultsFirstly, we found that seeding 10 × 103 cells at 70% cell confluency with 300 microcarriers per construct resulted in substantial increase in cell growth (76.8-fold increase in DNA) and chondrogenic protein generation (78.3- and 686-fold increase in GAG and Collagen II, respectively). Reducing cell density by adding empty microcarriers at seeding and indirectly compacting constructs by applying centrifugation at seeding or agitation throughout differentiation caused reduced cell growth and chondrogenic differentiation. Secondly, we showed that cell attachment to microcarriers throughout differentiation improves cell growth and chondrogenic outcomes since critically defined heMSC-Cytodex 1 constructs developed larger diameters (2.6-fold), and produced more DNA (13.8-fold), GAG (11.0-fold), and Collagen II (6.6-fold) than their equivalent cell-only counterparts. Thirdly, heMSC-Cytodex 1/3 constructs generated with cell-laden microcarriers from 1-day attachment in shake flask cultures were more efficient than those from 5-day expansion in spinner cultures in promoting cell growth and chondrogenic output per construct and per cell. Lastly, we demonstrate that these critically defined parameters can be applied across multiple microcarrier types, such as Cytodex 3, SphereCol and Cultispher-S, achieving similar trends in enhancing cell growth and chondrogenic differentiation.ConclusionsThis is the first study that has identified a set of critical attributes that enables efficient chondrogenic differentiation of heMSC-microcarrier constructs across multiple microcarrier types. It is also the first to demonstrate that cell attachment to microcarriers throughout differentiation improves cell growth and chondrogenic outcomes across different microcarrier types, including biodegradable gelatin-based microcarriers, making heMSC-microcarrier constructs applicable for use in allogeneic cartilage cell therapy.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0538-x) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 90%

Critical attributes of human early mesenchymal stromal cell-laden microcarrier constructs for improved chondrogenic differentiation

et al. 2017

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“…Biomaterials also have been investigated for their potential role in improving BM-MSC proliferation and increasing the efficacy of their transplantation. Nanofibrous, ECM-based structures and engineered delivery systems, such as biodegradable polycaprolactone microcarriers, may encourage the expansion and engraftment of BM-MSCs, suggesting biomimetic BM-MSC-designed niches represent an appealing adjunctive strategy to encourage both BM-MSC proliferation and BM-MSC-based therapy (Yan et al, 2015; Bhardwaj and Webster, 2016; Shekaran et al 2016). …”

Section: Identifying the Optimal Cell Type For Stem Cell Transplanmentioning

confidence: 99%

Stem cell therapy for abrogating stroke-induced neuroinflammation and relevant secondary cell death mechanisms

Stonesifer

Corey

Ghanekar

et al. 2017

Progress in Neurobiology

198

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Ischemic stroke is a leading cause of death worldwide. A key secondary cell death mechanism mediating neurological damage following the initial episode of ischemic stroke is the upregulation of endogenous neuroinflammatory processes to levels that destroy hypoxic tissue local to the area of insult, induce apoptosis, and initiate a feedback loop of inflammatory cascades that can expand the region of damage. Stem cell therapy has emerged as an experimental treatment for stroke, and accumulating evidence supports the therapeutic efficacy of stem cells to abrogate stroke-induced inflammation. In this review, we investigate clinically relevant stem cell types, such as hematopoietic stem cells (HSCs), mesenchymal stem cells (MSCs), endothelial progenitor cells (EPCs), very small embryonic-like stem cells (VSELs), neural stem cells (NSCs), extraembryonic stem cells, adipose tissue-derived stem cells, breast milk-derived stem cells, menstrual blood-derived stem cells, dental tissue-derived stem cells, induced pluripotent stem cells (iPSCs), teratocarcinoma-derived Ntera2/D1 neuron-like cells (NT2N), c-mycER(TAM) modified NSCs (CTX0E03), and notch-transfected mesenchymal stromal cells (SB623), comparing their potential efficacy to sequester stroke-induced neuroinflammation and their feasibility as translational clinical cell sources. To this end, we highlight that MSCs, with a proven track record of safety and efficacy as a transplantable cell for hematologic diseases, stand as an attractive cell type that confers superior anti-inflammatory effects in stroke both in vitro and in vivo. That stem cells can mount a robust anti-inflammatory action against stroke complements the regenerative processes of cell replacement and neurotrophic factor secretion conventionally ascribed to cell-based therapy in neurological disorders.

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“…When expanded under these conditions, MSC population doubling time (PDT) ranges from 40 to 120 h (Heathman et al, 2015;Lambrechts et al, 2016;Rafiq et al, 2013) (Figure S6). Notably, assessments of stemness in these studies were mostly limited to the minimal criteria established by the ISCT (Dominici et al, 2006;Heathman et al, 2015;Lambrechts et al, 2016;Rafiq et al, 2013;Shekaran et al, 2016). In contrast, using 2D monolayer culture, we show that MSC HS8 have PDT ranging from 40 to 80 h (mean = 60 h), a result closer to the PDT of fetal MSCs (43 h), a phenotypically younger and more naive population of cells (Shekaran et al, 2016) ( Figure S6).…”

Section: Discussionmentioning

confidence: 79%

Enhancing the Efficacy of Stem Cell Therapy with Glycosaminoglycans

et al. 2020

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Human mesenchymal stem cell (hMSC) therapy offers significant potential for osteochondral regeneration. Such applications require their ex vivo expansion in media frequently supplemented with fibroblast growth factor 2 (FGF2). Particular heparan sulfate (HS) fractions stabilize FGF2-FGF receptor complexes. We show that an FGF2-binding HS variant (HS8) accelerates the expansion of freshly isolated bone marrow hMSCs without compromising their naivety. Importantly, the repair of osteochondral defects in both rats and pigs is improved after treatment with HS8-supplemented hMSCs (MSC HS8), when assessed histologically, biomechanically, or by MRI. Thus, supplementing hMSC culture media with an HS variant that targets endogenously produced FGF2 allows the elimination of exogenous growth factors that may adversely affect their therapeutic potency.

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Biodegradable ECM-coated PCL microcarriers support scalable human early MSC expansion and in vivo bone formation

Cited by 60 publications

References 23 publications

Critical attributes of human early mesenchymal stromal cell-laden microcarrier constructs for improved chondrogenic differentiation

Critical attributes of human early mesenchymal stromal cell-laden microcarrier constructs for improved chondrogenic differentiation

Stem cell therapy for abrogating stroke-induced neuroinflammation and relevant secondary cell death mechanisms

Enhancing the Efficacy of Stem Cell Therapy with Glycosaminoglycans

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