CXCR4 expression by mesenchymal stromal cells is lost after use of enzymatic dissociation agents, but preserved by use of non-enzymatic methods

Pervin, Burcu; Aydın, Gülümser; Visser, Trudi P.; Uçkan-Çetinkaya, Duygu; Aerts‐Kaya, Fatima

doi:10.1007/s12185-020-03043-0

Cited by 5 publications

(11 citation statements)

References 20 publications

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“…[44][45][46][47] Furthermore, induction of CXCR4 expression with IL-1β has been shown to promote regeneration after DSS-induced colitis by enhancing homing of BM-MSCs to damaged intestinal mucosa, in comparison to unstimulated BM-MSCs. 48 We have now shown that the use of most enzymatic dissociation methods, including trypsin, may affect CXCR4 expression by both human 11 and murine BM-MSCs. However, the absence of CXCR4 expression on murine BM-MSCs did not affect the regenerative potential of the cells, even though homing may have been decreased.…”

Section: Discussionmentioning

confidence: 98%

“…17 However, we have previously shown that the use of trypsin, which is standardly used for collection of culture expanded human BM-MSCs for both research and clinical purposes, is correlated with a significant decrease in surface expression of CXCR4. 11 Although in vivo use of MSCs has shown a favorable safety profile, large discrepancies in the expected/predicted effects, based on in vitro data, 18,19 suggest that these differences in MSC potency may be related to the fact that the stem cells may not (or not in sufficient numbers) reach their target organ/tissue. Therefore, we hypothesized that the use of trypsin may cut the CXCL12 binding domain of CXCR4, effectively rendering the MSCs blind and unable to migrate to areas of injury in response to a CXCL12 gradient.…”

Section: Introductionmentioning

confidence: 99%

“…In addition, preparation of infusible cell concentrates requires the production of single‐cell suspensions via collection of the adherent cells by enzymatic dissociation solutions, such as trypsin with or without the addition of EDTA 17 . However, we have previously shown that the use of trypsin, which is standardly used for collection of culture expanded human BM‐MSCs for both research and clinical purposes, is correlated with a significant decrease in surface expression of CXCR4 11 . Although in vivo use of MSCs has shown a favorable safety profile, large discrepancies in the expected/predicted effects, based on in vitro data, 18,19 suggest that these differences in MSC potency may be related to the fact that the stem cells may not (or not in sufficient numbers) reach their target organ/tissue.…”

Section: Introductionmentioning

confidence: 99%

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Bone marrow mesenchymal stromal cells support regeneration of intestinal damage in a colitis mouse model, independent of their CXCR4 expression

Pervin,

Gizer,

Şeker

et al. 2024

Clinical Translational Sci

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Inflammatory bowel disease (IBD) is characterized by a chronically dysregulated immune response in the gastrointestinal tract. Bone marrow multipotent mesenchymal stromal cells have an important immunomodulatory function and support regeneration of inflamed tissue by secretion of soluble factors as well as through direct local differentiation. CXCR4 is the receptor for CXCL12 (SDF‐1, stromal‐derived factor‐1) and has been shown to be the main chemokine receptor, required for homing of MSCs. Increased expression of CXCL12 by inflamed intestinal tissue causes constitutive inflammation by attracting lymphocytes but can also be used to direct MSCs to sites of injury/inflammation. Trypsin is typically used to dissociate MSCs into single‐cell suspensions but has also been shown to digest surface CXCR4. Here, we assessed the regenerative effects of CXCR4high and CXCR4low MSCs in an immune‐deficient mouse model of DSS‐induced colitis. We found that transplantation of MSCs resulted in clinical improvement and histological recovery of intestinal epithelium. In contrary to our expectations, the levels of CXCR4 on transplanted MSCs did not affect their regenerative supporting potential, indicating that paracrine effects of MSCs may be largely responsible for their regenerative/protective effects.

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Section: Discussionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Bone marrow mesenchymal stromal cells support regeneration of intestinal damage in a colitis mouse model, independent of their CXCR4 expression

Pervin,

Gizer,

Şeker

et al. 2024

Clinical Translational Sci

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“…Therefore, behavioural activity after TFF2 injection with or without pre-infusion of CXCR4 is to be investigated as well [ 66 , 91 ]. Importantly, expression of CXCR4 in other tissues including digestive tract and metabolic tissues [ 92 , 93 , 94 , 95 , 96 , 97 , 98 , 99 , 100 ], could suggest a metabolic action of TFF2 via CXCR4 within these tissues.…”

Section: Tff2 Metabolic Properties and Implicationsmentioning

confidence: 99%

Trefoil Factor Family Member 2: From a High-Fat-Induced Gene to a Potential Obesity Therapy Target

Ghanemi

Yoshioka²,

St‐Amand

2021

Metabolites

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Obesity has its epidemiological patterns continuously increasing. With controlling both diet and exercise being the main approaches to manage the energy metabolism balance, a high-fat (HF) diet is of particular importance. Indeed, lipids have a low satiety potential but a high caloric density. Thus, focusing on pharmacologically targetable pathways remains an approach with promising therapeutic potential. Within this context, trefoil factor family member 2 (Tff2) has been characterized as specifically induced by HF diet rather than low-fat diet. TFF2 has also been linked to diverse neurological mechanisms and metabolic patterns suggesting its role in energy balance. The hypothesis is that TFF2 would be a HF diet-induced signal that regulates metabolism with a focus on lipids. Within this review, we put the spotlight on key findings highlighting this line of thought. Importantly, the hypothetical mechanisms pointed highlight TFF2 as an important contributor to obesity development via increasing lipids intestinal absorption and anabolism. Therefore, an outlook for future experimental activities and evaluation of the therapeutic potential of TFF2 inhibition is given. Indeed, its knockdown or downregulation would contribute to an antiobesity phenotype. We believe this work represents an addition to our understanding of the lipidic molecular implications in obesity, which will contribute to develop therapies aiming to manage the lipidic metabolic pathways including the absorption, storage and metabolism via targeting TFF2-related pathways. We briefly discuss important relevant concepts for both basic and clinical researchers.

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“…Enzymatic passages with dissociation protocols, in which trypsin with or without EDTA is typically used [ 1 , 2 ], are the most common methods employed to isolate the cells from confluent conditions and expand cultures. However, the dissociation using proteolytic enzymes may impair the viability and phenotype of the cells due to technical sensitivity [ 3 , 4 ]. Overtreating the cells with enzymatic digestions could degenerate proliferation capacity and influence their ability to differentiate into mature cells [ 5 ].…”

Section: Introductionmentioning

confidence: 99%

Serial Cultivation of an MSC-Like Cell Line with Enzyme-Free Passaging Using a Microporous Titanium Scaffold

et al. 2023

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In vitro studies on adherent cells require a process of passage to dissociate the cells from the culture substrate using enzymes or other chemical agents to maintain cellular activity. However, these proteolytic enzymes have a negative influence on the viability and phenotype of cells. The mesenchymal stem cell (MSC)-like cell line, C3H10T1/2, adhered, migrated, and proliferated to the same extent on newly designed microporous titanium (Ti) membrane and conventional culture dish, and spontaneous transfer to another substrate without enzymatic or chemical dissociation was achieved. The present study pierced a 10 μm-thick pure Ti sheet with 25 μm square holes at 75 μm intervals to create a dense porous structure with biomimetic topography. The pathway of machined holes allowed the cells to access both sides of the membrane frequently. In a culture with Ti membranes stacked above- and below-seeded cells, cell migration between the neighboring membranes was confirmed using the through-holes of the membrane and contact between the membranes as migration routes. Furthermore, the cells on each membrane migrated onto the conventional culture vessel. Therefore, a cell culture system with enzyme-free passaging was developed.

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CXCR4 expression by mesenchymal stromal cells is lost after use of enzymatic dissociation agents, but preserved by use of non-enzymatic methods

Cited by 5 publications

References 20 publications

Bone marrow mesenchymal stromal cells support regeneration of intestinal damage in a colitis mouse model, independent of their CXCR4 expression

Bone marrow mesenchymal stromal cells support regeneration of intestinal damage in a colitis mouse model, independent of their CXCR4 expression

Trefoil Factor Family Member 2: From a High-Fat-Induced Gene to a Potential Obesity Therapy Target

Serial Cultivation of an MSC-Like Cell Line with Enzyme-Free Passaging Using a Microporous Titanium Scaffold

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