Regulatory-compliant conditions during cell product manufacturing enhance in vitro immunomodulatory properties of infrapatellar fat pad-derived mesenchymal stem/stromal cells

ABSTRACTCD146+ bone marrow–derived Mesenchymal Stem/Stromal Cells (BM-MSC) play key roles in the perivascular niche, skeletogenesis and hematopoietic support, however elucidation of therapeutic potency has yet to be determined. Here, inflammatory challenge to crude BM-MSC captured a baseline of signatures including enriched expression of CD146+ with CD107a+, CXCR4+, and LepR+, transcriptional profile, enhanced secretory capacity, robust secretome and immunomodulatory function with stimulated target immune cells. These responses were significantly more pronounced in CD146+ (POS)-selected subpopulation than in the CD146- (NEG). Mechanistically, POS uniquely mediated robust immunosuppression while inducing significant frequencies of Naïve and Regulatory T cells in vitro. Moreover, POS promoted a pivotal M1-to-M2 macrophage shift in vivo, ameliorating inflammation/fibrosis of joint synovium and fat pad of the knee, failed by NEG. This study provides high-content evidence of CD146+CD107a+ BM-MSC, herein deemed ‘first responders’ to inflammation, as the underrepresented subpopulation within crude BM-MSC with innately higher secretory capacity and therapeutic potency.HIGHLIGHTSSignature phenotypic, transcriptional, and secretome profiles were identified and enriched in human CD146+ (POS)-selected subpopulation in response to inflammationInflammatory challenge consistently altered stemness (LIF) and differentiation master regulators (SOX9, RUNX2, PPARγ) in crude, POS, and NEG BM-MSC, and deduced unique expressions in POS compared to NEGPOS BM-MSC mediated the strongest immunomodulation, e.g. target immune cell suppression, Treg induction, diminished T cell differentiationPOS BM-MSC promoted the largest M1-to-M2 shift in vivo alleviating induced synovitis and infrapatellar fat pad fibrosis of the knee

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Infrapatellar fat pad-derived mesenchymal stromal cells from osteoarthritis patients: In vitro genetic stability and replicative senescence

Journal of Orthopaedic Research® ◽

10.1002/jor.23349 ◽

2016 ◽

Vol 35 (5) ◽

pp. 1029-1037 ◽

Cited By ~ 14

Author(s):

Simona Neri ◽

Serena Guidotti ◽

Nicoletta Libera Lilli ◽

Luca Cattini ◽

Erminia Mariani

Keyword(s):

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Genetic Stability ◽

Replicative Senescence ◽

Infrapatellar Fat Pad ◽

Fat Pad

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The clinical efficacy of arthroscopic therapy with knee infrapatellar fat pad cell concentrates in treating knee cartilage lesion: a prospective, randomized, and controlled study

Journal of Orthopaedic Surgery and Research ◽

10.1186/s13018-021-02224-9 ◽

2021 ◽

Vol 16 (1) ◽

Author(s):

Yiqin Zhou ◽

Haobo Li ◽

Dong Xiang ◽

Jiahua Shao ◽

Qiwei Fu ◽

...

Keyword(s):

Clinical Efficacy ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Knee Arthroscopy ◽

Infrapatellar Fat Pad ◽

Control Group ◽

Knee Cartilage ◽

Fat Pad ◽

Group 12 ◽

Significant Difference

Abstract Introduction To evaluate the clinical efficacy of arthroscopic therapy with infrapatellar fat pad cell concentrates in treating knee cartilage lesions, we conducted a prospective randomized single-blind clinical study of controlled method. Methods Sixty cases from Shanghai Changzheng Hospital from April 2018 to December 2019 were chosen and randomly divided into 2 groups equally. Patients in the experiment group were treated through knee arthroscopy with knee infrapatellar fat pad cell concentrates containing mesenchymal stromal cells, while patients in the control group were treated through regular knee arthroscopic therapy. VAS and WOMAC scores were assessed at pre-operation, and 6 weeks, 12 weeks, 6 months, and 12 months after intervention. MORCART scores were assessed at pre-operation and 12 months after intervention. Results Twenty-nine cases in the experiment group and 28 cases in the control group were followed up. No significant difference in VAS, WOMAC, and MOCART scores were found between the two groups before surgery (P > 0.05). The WOMAC total and WOMAC function scores of the experiment group were significantly lower than those of the control group 6 months and 12 months after surgery (P < 0.05). The VAS rest and VAS motion scores of the experiment group were found significantly lower than those of the control group 12 months after surgery (P < 0.05). The MOCART scores of the experiment group were found significantly higher compared with the control group 12 months after surgery (P < 0.05). No significant difference in WOMAC stiffness scores were found between the two groups. Conclusions The short-term results of our study are encouraging and demonstrate that knee arthroscopy with infrapatellar fat pad cell concentrates containing mesenchymal stromal cells is safe and provides assistance in reducing pain and improving function in patients with knee cartilage lesions. Trial registration ChiCTR1800015379. Registered on 27 March 2018, http://www.chictr.org.cn/showproj.aspx?proj=25901.

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Multipotent Stromal Cells Derived From the Infrapatellar Fat Pad of the Knee

Clinical Orthopaedics and Related Research ◽

10.1097/01.blo.0000072467.53786.ca ◽

2003 ◽

Vol 412 ◽

pp. 196-212 ◽

Cited By ~ 263

Author(s):

M. Quinn Wickham ◽

Geoffrey R. Erickson ◽

Jeffrey M. Gimble ◽

T. Parker Vail ◽

Farshid Guilak

Keyword(s):

Stromal Cells ◽

Infrapatellar Fat Pad ◽

Multipotent Stromal Cells ◽

Fat Pad

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Infrapatellar fat pad-derived mesenchymal stromal cell product for treatment of knee osteoarthritis: a first-in-human study with evaluation of the potency marker

Cytotherapy ◽

10.1016/j.jcyt.2021.08.006 ◽

2021 ◽

pp. 1377

Author(s):

Hung-Hsuan Chen ◽

Yu-Chun Chen ◽

San-Ni Yu ◽

Wan-Ling Lai ◽

Yi-Shan Shen ◽

...

Keyword(s):

Knee Osteoarthritis ◽

Mesenchymal Stromal Cell ◽

Stromal Cell ◽

Human Study ◽

Infrapatellar Fat Pad ◽

Fat Pad ◽

Cell Product

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Effect of extracorporeal shock wave therapy on equine umbilical cord blood mesenchymal stromal cells in vitro

10.1101/2020.01.10.901439 ◽

2020 ◽

Author(s):

Ramés Salcedo-Jiménez ◽

Judith Koenig ◽

Olivia Lee ◽

Thomas W.G. Gibson ◽

Pavneesh Madan ◽

...

Keyword(s):

Shock Wave ◽

Mesenchymal Stromal Cells ◽

Umbilical Cord Blood ◽

Stromal Cells ◽

Extracorporeal Shock Wave Therapy ◽

Shock Wave Therapy ◽

Extracorporeal Shock Wave ◽

Immunomodulatory Properties

AbstractExtracorporeal shock wave therapy (ESWT) has been shown to induce different biological effects on a variety of cells, including regulation and stimulation of their function and metabolism. ESWT can promote different biological responses such as proliferation, migration, and regenerations of cells. Recent studies have shown that mesenchymal stromal cells (MSCs) secrete factors that enhance the regeneration of tissues, stimulate proliferation and differentiation of cells and decrease inflammatory and immune-reactions. Clinically, the combination of these two therapies has been used as a treatment for tendon and ligament lesions in horses; however, there is no scientific evidence supporting this combination of therapies in vivo. Therefore, the objectives of the study were to evaluate the effects of ESWT on equine umbilical cord blood mesenchymal stromal cells (CB-MSCs) proliferative, metabolic, migrative, differentiation, and immunomodulatory properties in vitro. Three equine CB-MSC cultures from independent donors were treated using an electrohydraulic shock wave generator attached to a water bath. All experiments were performed as triplicates. Proliferation, viability, migration and immunomodulatory properties of the cells were evaluated. Equine CB-MSCs were induced to evaluate their trilineage differentiation potential. ESWT treated cells had increased metabolic activity, showed positive adipogenic, osteogenic, and chondrogenic differentiation, and showed higher potential for differentiation towards the adipogenic and osteogenic cell fates. ESWT treated cells showed similar immunomodulatory properties to none-ESWT treated cells. Equine CB-MSCs are responsive to ESWT treatment and showed increased metabolic, adipogenic and osteogenic activity, but unaltered immunosuppressive properties. In vivo studies are warranted to determine if synergistic effects occur in the treatment of musculoskeletal injuries if ESWT and equine CB-MSC therapies are combined.

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Infrapatellar Fat Pad/Synovium Complex in Early-Stage Knee Osteoarthritis: Potential New Target and Source of Therapeutic Mesenchymal Stem/Stromal Cells

Frontiers in Bioengineering and Biotechnology ◽

10.3389/fbioe.2020.00860 ◽

2020 ◽

Vol 8 ◽

Cited By ~ 3

Author(s):

Dylan N. Greif ◽

Dimitrios Kouroupis ◽

Christopher J. Murdock ◽

Anthony J. Griswold ◽

Lee D. Kaplan ◽

...

Keyword(s):

Knee Osteoarthritis ◽

Stromal Cells ◽

Early Stage ◽

Infrapatellar Fat Pad ◽

Fat Pad

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Human Infrapatellar Fat Pad-Derived Stromal Cells have more Potent Differentiation Capacity than other Mesenchymal Cells and can be Enhanced by Hyaluronan

Cell Transplantation ◽

10.3727/096368914x681937 ◽

2015 ◽

Vol 24 (7) ◽

pp. 1221-1232 ◽

Cited By ~ 19

Author(s):

Dah-Ching Ding ◽

Kun-Chi Wu ◽

Hsiang-Lan Chou ◽

Wei-Ting Hung ◽

Hwan-Wun Liu ◽

...

Keyword(s):

Stromal Cells ◽

Mesenchymal Cells ◽

Infrapatellar Fat Pad ◽

Fat Pad ◽

Differentiation Capacity

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Interferon-Gamma Impairs Expansion and Hematopoietic Support of Bone Marrow Mesenchymal Stromal Cells

Blood ◽

10.1182/blood.v128.22.3884.3884 ◽

2016 ◽

Vol 128 (22) ◽

pp. 3884-3884

Author(s):

Marieke Goedhart ◽

Anne Cornelissen ◽

Carlijn Kuijk ◽

Sulima Geerman ◽

Fernanda Pascutti ◽

...

Keyword(s):

Stromal Cells ◽

Stromal Compartment ◽

Self Renewal ◽

Hematopoietic Stem ◽

Ifn Γ ◽

Immunomodulatory Properties ◽

Pre Treatment ◽

The Impact

Abstract Maintenance of hematopoietic stem cells (HSCs) and regulation of their quiescence and self-renewal is critical for maintaining a lifelong supply of blood cells. The ability of HSCs to stay quiescent is thought to depend on their specific niche in the bone marrow (BM). Mesenchymal stromal cells (MSC) in the BM are multipotent stem cells that form part of the vascular HSC niche and provide micro-environmental support to HSCs both in vivo and upon expansion ex vivo. Culture-expanded MSCs also exhibit immunomodulatory properties that can be enhanced by pre-treatment with interferon-gamma (IFN-γ). BM MSC are thus attractive candidates for cellular therapy after hematopoietic stem cell transplantation, for promoting rapid hematopoietic recovery and reducing the incidence or severity of graft versus host disease. Although IFN-γ pre-treatment can improve the immunomodulatory properties of MSCs, elevated IFN-γ levels have also been associated with anemia and BM failure in multiple chronic inflammatory diseases. While the impact of IFN-γ on HSC has been elucidated in recent years, it remains largely unknown whether IFN-γ can also influence hematopoietic support by BM stromal cells. In this study, we aim to elucidate the impact of IFN-γ on hematopoietic support of BM MSC. We show that in vitro expansion of primary BM MSC cultures from healthy donors was significantly reduced in the presence of IFN-γ, and this effect could be reproduced in the BM stromal cell line MS-5. Concurrently, IFN-γ diminished the clonal capacity of BM MSC, as measured by CFU-F assays. In addition, BM MSC that were pre-stimulated with IFN-γ produced significantly lower levels of CXCL12, suggesting a loss of hematopoietic support potential. Indeed, support of CD34+ hematopoietic stem and progenitor cells (HSPC) in a co-culture assay was greatly reduced in when MSC were pre-treated with IFN-γ. To determine the impact of IFN-γ on BM MSC in vivo, we investigated the BM stromal compartment of IFN-γ AU-rich element deleted (ARE-Del) mice, which constitutively express IFN-γ in steady state conditions. FACS analysis revealed a remodeling of the BM stromal compartment in ARE-Del mice compared to littermate controls, with significantly fewer MSCs, identified as CD45-Ter119-CD31-CD51+PDGFRa+ cells. Numbers of other stromal cell subsets, such as osteoblasts and fibroblasts, were not altered. The reduction of BM MSC in ARE-Del mice coincided with a loss of quiescence in HSCs; only 35% of long term HSC (LT-HSC) in ARE-Del mice were quiescent, compared to 70% in WT mice, as determined by Ki-67 staining. Loss of quiescence in LT-HSC did not lead to increased self-renewal, but rather induced increased differentiation towards short-term HSC and multi-potent progenitors. We then sorted LT-HSC from WT and ARE-Del mice and performed in vitro HSC culture assays in the absence of IFN-γ. Absolute numbers of LT-HSC were rapidly decreased in ARE-Del compared to WT cultures after 3 and 7 days of HSC culture, while numbers of more differentiated progenitors were increased. These data indicate that an IFN-γ-mediated loss of BM MSC in ARE-Del mice leads to loss of quiescent LT-HSCs and induces a tendency towards HSC differentiation over self-renewal. In conclusion, we have shown that IFN-γ has a negative impact on expansion and hematopoietic support of BM MSC in vitro and in vivo across species. Although IFN-γ treatment enhances the immunomodulatory function of MSCs in a clinical setting, it is obvious from our data that IFN-γ impairs their HSC supporting function. These data also provide more insight in the underlying mechanism by which IFN-γ contributes to the pathogenesis of anemia and BM failure. Disclosures No relevant conflicts of interest to declare.

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