Development of Novel Monoclonal Antibodies that Define Differentiation Stages of Human Stromal (Mesenchymal) Stem Cells

Andersen, Ditte Caroline; Kortesidis, Angela; Zannettino, Andrew C.W.; Kratchmarova, Irina; Chen, Li; Jensen, Ole Nørregaard; Teisner, B.; Gronthos, Stan; Jensen, Charlotte Harken; Kassem, Moustapha

doi:10.1007/s10059-011-2277-7

Cited by 14 publications

(9 citation statements)

References 36 publications

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“…These observations impair its clinical use by increasing the likelihood of variation to the extent of bone healing. More recently, collagen IV was shown to select for cultured, but undifferentiated MSCs [55]. In contrast, we show that continuous supplementation with HS-2 similarly upregulates hMSC self-renewal and greatly improves their homogeneity, suggesting that HS-2 would benefit future therapies utilizing cultured hMSCs.…”

Section: Discussionmentioning

confidence: 43%

Heparan Sulfate Enhances the Self-Renewal and Therapeutic Potential of Mesenchymal Stem Cells from Human Adult Bone Marrow

Helledie

Dombrowski

Rai

et al. 2012

Stem Cells and Development

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Insufficient cell number hampers therapies utilizing adult human mesenchymal stem cells (hMSCs) and current ex vivo expansion strategies lead to a loss of multipotentiality. Here we show that supplementation with an embryonic form of heparan sulfate (HS-2) can both increase the initial recovery of hMSCs from bone marrow aspirates and increase their ex vivo expansion by up to 13-fold. HS-2 acts to amplify a subpopulation of hMSCs harboring longer telomeres and increased expression of the MSC surface marker stromal precursor antigen-1. Gene expression profiling revealed that hMSCs cultured in HS-2 possess a distinct signature that reflects their enhanced multipotentiality and improved bone-forming ability when transplanted into critical-sized bone defects. Thus, HS-2 offers a novel means for decreasing the expansion time necessary for obtaining therapeutic numbers of multipotent hMSCs without the addition of exogenous growth factors that compromise stem cell fate.

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Section: Discussionmentioning

confidence: 43%

Heparan Sulfate Enhances the Self-Renewal and Therapeutic Potential of Mesenchymal Stem Cells from Human Adult Bone Marrow

Helledie

Dombrowski

Rai

et al. 2012

Stem Cells and Development

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“…In undifferentiated cells, type VI collagen was localized exclusively to the cytoplasm. In contrast, when the media were changed to stimulate osteogenesis or adipogenesis, type VI collagen was found localized to the cell membrane and pericellular matrix [50]. More recently, Urciuolo et al (2013) demonstrated that type VI collagen plays a key role in creating the muscle satellite cell niche.…”

Section: Discussionmentioning

confidence: 99%

One size does not fit all: developing a cell-specific niche for in vitro study of cell behavior

Marinkovic¹,

Block²,

Rakian

et al. 2016

Matrix Biology

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For more than 100 years, cells and tissues have been studied in vitro using glass and plastic surfaces. Over the last 10–20 years, a great body of research has shown that cells are acutely sensitive to their local environment (extracellular matrix, ECM) which contains both chemical and physical cues that influence cell behavior. These observations suggest that modern cell culture systems, using tissue culture polystyrene (TCP) surfaces, may fail to reproduce authentic cell behavior in vitro, resulting in “artificial outcomes.” In the current study, we use bone marrow (BM)- and adipose (AD)-derived stromal cells to prepare BM-ECM and AD-ECM, which are decellularized after synthesis by the cells, to mimic the cellular niche for each of these tissues. Each ECM was characterized for its ability to affect BM- and AD-mesenchymal stem cell (MSC) proliferation, as well as proliferation of three cancer cell lines (HeLa, MCF-7, and MDA-MB-231), modulate cell spreading, and direct differentiation relative to standard TCP surfaces. We found that both ECMs promoted the proliferation of MSCs, but that this effect was enhanced when the tissue-origin of the cells matched that of the ECM (i.e. BM-ECM promoted the proliferation of BM-MSCs over AD-MSCs, and vice versa). Moreover, BM- and AD-ECM were shown to preferentially direct MSC differentiation towards either osteogenic or adipogenic lineage, respectively, suggesting that the effects of the ECM were tissue-specific. Further, each ECM influenced cell morphology (i.e. circularity), irrespective of the origin of the MSCs, lending more support to the idea that effects were tissue specific. Interestingly, unlike MSCs, these ECMs did not promote the proliferation of the cancer cells. In an effort to further understand how these three culture substrates influence cell behavior, we evaluated the chemical (protein composition) and physical properties (architecture and mechanical) of the two ECMs. While many structural proteins (e.g. collagen and fibronectin) were found at equivalent levels in both BM- and AD-ECM, the architecture (i.e. fiber orientation; surface roughness) and physical properties (storage modulus, surface energy) of each were unique. These results, demonstrating differences in cell behavior when cultured on the three different substrates (BM- and AD-ECM and TCP) with differences in chemical and physical properties, provide evidence that the two ECMs may recapitulate specific elements of the native stem cell niche for bone marrow and adipose tissues. More broadly, it could be argued that ECMs, elaborated by cells ex vivo, serve as an ideal starting point for developing tissue-specific culture environments. In contrast to TCP, which relies on the “one size fits all” paradigm, native tissue-specific ECM may be a more rational model to approach engineering 3D tissue-specific culture systems to replicate the in vivo niche. We suggest that this approach will provide more meaningful information for basic research studies of cell behavior as well as cell-based therapeutics.

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“…For direct ELISAs, purified mouse DLK1 (0.5-1.0 µg/mL) was coated onto 96-well Maxisorp flat bottom microtiter plates. For cyto-ELISAs, 3T3-L1 and C2C12 cells were cultured in 96-well plates and fixed with 4% NBF at sub-confluence, as previously described [22]. Hybridoma supernatants were then transferred to the wells and plates incubated overnight.…”

Section: Screening Of Hybridoma Supernatantsmentioning

confidence: 99%

Antibody-based inhibition of circulating DLK1 protects from estrogen deficiency-induced bone loss in mice

Figeac

Andersen

Nielsen

et al. 2018

Bone

Self Cite

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Soluble delta-like 1 homolog (DLK1) is a circulating protein that belongs to the Notch/Serrate/delta family, which regulates many differentiation processes including osteogenesis and adipogenesis. We have previously demonstrated an inhibitory effect of DLK1 on bone mass via stimulation of bone resorption and inhibition of bone formation. Further, serum DLK1 levels are elevated and positively correlated to bone turnover markers in estrogen (E)-deficient rodents and women. In this report, we examined whether inhibition of serum DLK1 activity using a neutralizing monoclonal antibody protects from E deficiency-associated bone loss in mice. Thus, we generated mouse monoclonal anti-mouse DLK1 antibodies (MAb DLK1) that enabled us to reduce and also quantitate the levels of bioavailable serum DLK1 in vivo. Ovariectomized (ovx) mice were injected intraperitoneally twice weekly with MAb DLK1 over a period of one month. DEXA-, microCT scanning, and bone histomorphometric analyses were performed. Compared to controls, MAb DLK1 treated ovx mice were protected against ovx-induced bone loss, as revealed by significantly increased total bone mass (BMD) due to increased trabecular bone volume fraction (BV/TV) and inhibition of bone resorption. No significant changes were observed in total fat mass or in the number of bone marrow adipocytes. These results support the potential use of anti-DLK1 antibody therapy as a novel intervention to protect from E deficiency associated bone loss.

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Development of Novel Monoclonal Antibodies that Define Differentiation Stages of Human Stromal (Mesenchymal) Stem Cells

Cited by 14 publications

References 36 publications

Heparan Sulfate Enhances the Self-Renewal and Therapeutic Potential of Mesenchymal Stem Cells from Human Adult Bone Marrow

Heparan Sulfate Enhances the Self-Renewal and Therapeutic Potential of Mesenchymal Stem Cells from Human Adult Bone Marrow

One size does not fit all: developing a cell-specific niche for in vitro study of cell behavior

Antibody-based inhibition of circulating DLK1 protects from estrogen deficiency-induced bone loss in mice

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