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cell transplantation for the treatment of spinal cord injury–bone marrow stromal cells and choroid plexus epithelial cells

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Neural Regeneration Research 2016年第9期11卷 1385-1388页

作者： Chizuka Ide Norihiko Nakano Kenji Kanekiyo Central Research Laboratory Aino University School of Health Science

Transplantation of bone marrow stromal cells （BMSCs） enhanced the outgrowth of regenerating axons and promoted locomotor improvements of rats with spinal cord injury （SCI）. BMSCs did not survive long-term, disappearing from the spinal cord within 2-3 weeks after transplantation. Astrocyte-devoid areas, in which no astrocytes or oligodendrocytes were found, formed at the epicenter of the lesion. It was remarkable that numerous regenerating axons extended through such astrocyte-devoid areas. Regenerating axons were associated with Schwann cells embedded in extracellular matrices. Transplantation of choroid plexus epithelial cells （CPECs） also enhanced axonal regeneration and locomotor improvements in rats with SCI. Although CPECs disappeared from the spinal cord shortly after transplantation, an extensive outgrowth of regenerating axons occurred through astrocyte-devoid areas, as in the case of BMSC transplantation. These findings suggest that BMSCs and CPECs secret neurotrophic factors that promote tissue repair of the spinal cord, including axonal regeneration and reduced cavity formation. This means that transplantation of BMSCs and CPECs promotes ＂intrinsic＂ ability of the spinal cord to regenerate. The treatment to stimu- late the intrinsic regeneration ability of the spinal cord is the safest method of clinical application for SCI. It should be emphasized that the generally anticipated long-term survival, proliferation and differentiation of transplanted cells are not necessarily desirable from the clinical point of view of safety.

关键词： bone marrow stromal cell choroid plexus epithelial cell spinal cord injury axonal regeneration locomotor improvement intrinsic regeneration ability

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3D printing of osteocytic Dll4 integrated with PCL for cell fate determination towards osteoblasts in vitro

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Bio-Design and Manufacturing 2022年第3期5卷 497-511页

作者： Pengtao Wang Xiaofang Wang Bo Wang Xian Li Zhengsong Xie Jie Chen Tasuku Honjo Xiaolin Tu Laboratory of Skeletal Development and Regeneration Institute of Life SciencesChongqing Medical UniversityChongqing 400016China Medical Data Science Academy Chongqing Medical UniversityChongqing 400016China Department of Immunology and Genomic Medicine Graduate School of MedicineKyoto UniversityYoshida Konoe-choSakyo-kuKyoto 606-8501Japan

Since 3D printed hard materials could match the shape of bone,cell survival and fate determination towards osteoblasts in such materials have become a popular research target.In this study,a scaffold of hardmaterial for 3D fabrication was designed to regulate developmental signal(Notch)transduction guiding osteoblast differentiation.We established a polycaprolactone(PCL)and cell-integrated 3D printing system(PCI3D)to reciprocally print the beams of PCL and cell-laden hydrogel for a module.This PCI3D module holds good cell viability of over 87%,whereas cells show about sixfold proliferation in a 7-day culture.The osteocytic MLO-Y4 was engineered to overexpress Notch ligand Dll4,making up 25%after mixing with 75%stromal cells in the PCI3D module.Osteocytic Dll4,unlike other delta-like family members such as Dll1 or Dll3,promotes osteoblast differentiation and themineralization of primary mouse and a cell line of bone marrow stromal cells when cultured in a PCI3D module for up to 28 days.Mechanistically,osteocytic Dll4 could not promote osteogenic differentiation of the primary bone marrow stromal cells(BMSCs)after conditional deletion of the Notch transcription factor RBPjκby Cre recombinase.These data indicate that osteocytic Dll4 activates RBPjκ-dependent canonical Notch signaling in BMSCs for their oriented differentiation towards osteoblasts.Additionally,osteocytic Dll4 holds a great potential for angiogenesis in human umbilical vein endothelial cells within modules.Our study reveals that osteocytic Dll4 could be the osteogenic niche determining cell fate towards osteoblasts.This will open a new avenue to overcome the current limitation of poor cell viability and low bioactivity of traditional orthopedic implants.

关键词： Integrated 3D printing PCL scaffold cell-laden hydrogel Osteocytic Dll4 bone marrow stromal cell Osteoblast differentiation cell viability in hard material RBPjκ Notch signaling

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bone marrow stromal cells Express Neural Phenotypes in vitro and Migrate in Brain After Transplantation in vivo

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Biomedical and Environmental Sciences 2006年第5期19卷 329-335页

作者： LI-YE YAN TIAN-HUA HUANG LIAN MA Department of Biology Medical College of Shantou University Shantou 515031 Guangdong China

Objective To investigate the differentiation of bone marrow stromal cells （BMSC） into neuron-like cells and to explore their potential use for neural transplantation. Methods BMSC from rats and adult humans were cultured in serum-containing media. Salvia miltiorrhiza was used to induce human BMSC （hBMSC） to differentiate. BMSC were identified with immunocytochemistry. Semi-quantitative RT-PCR was used to examine mRNA expression of neurofilamentl （NF1）, nestin and neuron-specific enolase （NSE） in rat BMSC （rBMSC）. Rat BMSC labelled by Hoschst33258 were transplanted into striatum of rats to trace migration and distribution. Results rBMSC expressed NSE, NFI and nestin mRNA, and NF1 mRNA and expression was increased with induction of Salvia miltiorrhiza. A small number of hBMSC were stained by anti-nestin, anti-GFAP and anti-S 100. Salvia miltiorrhiza could induce hBMSC to differentiate into neuron-like cells. Some differentiated neuron-like cells, that expressed NSE, beta-tubulin and NF-200, showed typical neuron morphology, but some neuron-like cells also expressed alpha smooth muscle protein, making their neuron identification complicated, rBMSC could migrate and adapted in the host brains after being transplanted. Conclusion bone marrow stromal cells could express phenotypes of neurons, and Salvia milliorrhiza could induce hBMSC to differentiate into neuron-like cells, If BMSC could be converted into neurons instead of mesenchymal derivatives, they would be an abundant and accessible cellular source to treat a variety of neurological diseases.

关键词： bone marrow stromal cell cell transplantation Differentiation Neuron Stem cell Salvia miltiorrhiza

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bone marrow stromal cells as a therapeutic treatment for ischemic stroke

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Neuroscience Bulletin 2014年第3期30卷 524-534页

作者： Zizhen Yang Lei Zhu Fangqin Li Jing Wang Huan Wan Yujun Pan Department of Neurology First Hospital and Clinical College Harbin Medical University

Cerebral ischemia remains the most frequent cause of death and quality-of-life impairments due to neurological deficits, and accounts for the majority of total healthcare costs. However, treatments for cerebral ischemia are limited. Over the last decade, bone marrow stromal cell （BMSC） therapy has emerged as a particularly appealing option, as it is possible to help patients even when initiated days or even weeks after the ischemic insult. BMSCs are a class of multipotent, self-renewing cells that give rise to differentiated progeny when implanted into appropriate tissues. Therapeutic effects of BMSC treatment for ischemic stroke, including sensory and motor recovery, have been reported in pre-clinical studies and clinical trials. In this article, we review the recent progress in BMSC-based therapy for ischemic stroke, focusing on the route of delivery and pre-processing of BMSCs. Selecting an optimal delivery route is of particular importance. The ideal approach, as well as the least risky, for translational applications still requires further identification. Appropriate pre- processing of BMSCs or combination therapy has the benefit of achieving the maximum possible restoration. Further pre-clinical studies are required to determine the time-window for transplantation and the appropriate dosage of cells.

关键词： bone marrow stromal cell cerebral ischemia transplantation neuroprotection administration method

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Effect of zinc substitution in hydroxyapatite coating on osteoblast and osteoclast differentiation under osteoblast/osteoclast co-culture

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Regenerative Biomaterials 2019年第6期6卷 349-359页

作者： Guolong Meng Xiaoli Wu Ruijuan Yao Jing He Wu Yao Fang Wu National Engineering Research Center for Biomaterials Sichuan UniversityChengdu 610064P.R.China

Zinc is an essential trace element required for bone remodelling process,but its role in such process remains to be elucidated.In particular,inconsistent results have been reported on the effect of Zn on osteoclastic responses,and supplement of receptor activator of nuclear factor kappa-B ligand(RANKL)factors has been commonly adopted.Co-culture is a suitable approach to elucidating the role of Zn in bone remodelling process,by better imitating the cellular environment as the presence of osteoblasts plays critical role in modulating osteoclastic functions.In this study,zincsubstituted HA coatings have been deposited using a liquid precursor plasma spraying process at two different concentrations(1,2 wt.%).The effect of zinc substitution on osteoblastic and osteoclastic differentiation has been studied in vitro.In particular,a cultivation regime was designed to first induce osteoblastic differentiation of rat bone marrow stromal cells(BMSCs)for 14 days,and then induce osteoclastic differentiation of osteoclast-like precursor RAW 264.7 cells through the aid of the osteoblasts formed for additional 14 days,in the absence of the external addition of RANKL.The results showed that Zn substitution moderately promoted the BMSC differentiation into the osteoblasts and reduced the osteoclastic activity in early time(1 day co-culture).However,promotion of the osteoclastic activity were observed at later stages,as indicated by the significantly enhanced expressions of trap5b and IL-1(8-and 15-day co-culture)and moderate stimulation of the nucleus integration and formation of the multinucleated cells(14-day co-culture).Such stimulating effect of the osteoclastic activity was absent under mono-culture of RAW 264.7 cell,with simple RANKL supplementation.The results suggest that both the zinc and the presence of MSC/osteoblast play profound and highly interacted roles on osteoclast differentiation and activity,which is critical in modulating the bone remodelling process.

关键词： zinc bone marrow stromal cell osteoclast precursor osteoclast co-cultivation bone remodelling process

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Role of endogenous Schwann cells in tissue repair after spinal cord injury

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Neural Regeneration Research 2013年第2期8卷 177-185页

作者： Shu-xin Zhang Fengfa Huang Mary Gates Eric G. Holmberg Spinal Cord Society Research Center Department of Chemistry University of Alaska

Schwann cells are glial cells of peripheral nervous system, responsible for axonal myelination and ensheathing, as well as tissue repair following a peripheral nervous system injury. They are one of several cell types that are widely studied and most commonly used for cell transplantation to treat spinal cord injury, due to their intrinsic characteristics including the ability to secrete a variety of neurotrophic factors. This mini review summarizes the recent findings of endogenous Schwann cells after spinal cord injury and discusses their role in tissue repair and axonal regeneration. After spinal cord injury, numerous endogenous Schwann cells migrate into the lesion site from the nerve roots, involving in the construction of newly formed repaired tissue and axonal myelination. These invading Schwann cells also can move a long distance away from the injury site both rostrally and caudally. In addition, Schwann cells can be induced to migrate by minimal insults （such as scar ablation） within the spinal cord and integrate with astrocytes under certain circumstances. More importantly, the host Schwann cells can be induced to migrate into spinal cord by transplantation of different cell types, such as exogenous Schwann cells, olfactory ensheathing cells, and bone marrow-derived stromal stem cells. Migration of endogenous Schwann cells following spinal cord injury is a common natural phenomenon found both in animal and human, and the myelination by Schwann cells has been examined effective in signal conduction electrophysiologically. Therefore, if the inherent properties of endogenous Schwann cells could be developed and utilized, it would offer a new avenue for the restoration of injured spinal cord.

关键词： neural regeneration spinal cord injury Schwann cells spinal cord injury tissue repair axonalregeneration myelination rat scar ablation astrocytes cell transplantation rose Bengal olfactoryensheathing cells bone marrow stromal cell grant-supported paper neuroregeneration

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Transcriptomic alterations underline aging of osteogenic bone marrow stromal cells

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World Journal of Stem cells 2021年第1期13卷 128-138页

作者： Yu-Hao Cheng Shu-Fen Liu Jing-Cheng Dong Qin Bian Institute for cell Engineering Johns Hopkins University School of MedicineBaltimoreMD 21205United States Institute of Spine Shanghai University of Traditional Chinese MedicineShanghai 200030China Department of Integrative Medicine Huashan HospitalFudan UniversityShanghai 200040China

BACKGROUND Multipotent bone marrow stromal cells(BMSCs)are adult stem cells that form functional osteoblasts and play a critical role in bone remodeling.During aging,an increase in bone loss and reduction in structural integrity lead to osteoporosis and result in an increased risk of fracture.We examined age-dependent histological changes in murine vertebrae and uncovered that bone loss begins as early as the age of 1 mo.AIM To identify the functional alterations and transcriptomic dynamics of BMSCs during early bone loss.METHODS We collected BMSCs from mice at early to middle ages and compared their selfrenewal and differentiation potential.Subsequently,we obtained the transcriptomic profiles of BMSCs at 1 mo,3 mo,and 7 mo.RESULTS The colony-forming and osteogenic commitment capacity showed a comparable finding that decreased at the age of 1 mo.The transcriptomic analysis showed the enrichment of osteoblastic regulation genes at 1 mo and loss of osteogenic features at 3 mo.The BMSCs at 7 mo showed enrichment of adipogenic and DNA repair features.Moreover,we demonstrated that the WNT and MAPK signaling pathways were upregulated at 1 mo,followed by increased pro-inflammatory and apoptotic features.CONCLUSION Our study uncovered the cellular and molecular dynamics of bone aging in mice and demonstrated the contribution of BMSCs to the early stage of age-related bone loss.

关键词： bone marrow stromal cell Mesenchymal stem cell Mesenchymal stromal cell Aging bone modeling and remodeling Transcriptome

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Intra-portal transplantation of bone marrow stromal cells ameliorates liver fibrosis in mice

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Hepatobiliary & Pancreatic Diseases International 2008年第3期7卷 264-270页

作者： Zheng, Jin-Fang Liang, Li-Jian Peoples Hosp Hainan Prov Dept Hepatobiliary Surg Haikou 570311 Peoples R China Sun Yat Sen Univ Affiliated Hosp 1 Dept Hepatobiliary Surg Guangzhou 510080 Peoples R China

BACKGROUND: bone marrow cells can differentiate into hepatocytes in a suitable microenvironment. This study was undertaken to investigate the effects of transplanted bone marrow stromal cells (BMSCs) on liver fibrosis in mice. METHODS: BMSCs were harvested and cultured from male BALB/c mice, then transplanted into female syngenic BALB/c mice via the portal vein. After partial hepatectomy, diethylnitrosamine (DEN) was administered to induce liver fibrosis. Controls received BMSCs and non-supplemented drinking water, the model group received DEN with their water, and the experimental group received BMSCs and DEN. Mice were killed after 3 months, and ALT, AST, hyaluronic acid (HA), and laminin (LN) in serum and hydroxyproline (Hyp) in the liver were assessed. Alpha-smooth muscle actin (alpha-SMA) in the liver was assessed by immunohistochemistry. bone marrow-derived hepatocytes were identified by fluorescent in situ hybridization (FISH) in liver sections. RESULTS: BMSCs were shown to differentiate into hepatocyte-like phenotypes after hepatocyte growth factor treatment in vitro. Serum ALT, AST, HA, and LN were markedly reduced by transplanted BMSCs. Liver Hyp content and alpha-SMA staining in mice receiving BMSCs were lower than in the model group, consistent with altered liver pathology. FISH analysis revealed the presence of donor-derived hepatocytes in the injured liver after cross-gender mouse BMSC transplantation. After three months, about 10% of cells in the injured liver were bone marrow-derived. CONCLUSION: BMSCs transplanted via the portal vein can convert into hepatocytes to repair liver injury induced by DEN, restore liver function, and reduce liver fibrosis.

关键词： bone marrow stromal cell hepatocyte differentiation cell therapy liver fibrosis

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Experimental Study on Allogenic Decalcified bone Matrix as Carrier for bone Tissue Engineering

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2004年第2期24卷 147-150页

作者：郑东杨述华李进许伟华杨操刘勇潘海涛黄自锋 Department of Orthopedics Union Hospital Tongji Medical College Huazhong University of Science and Technology

The biocompatibility and osteogenic activity of allogenic decalcified bone matrix (DBM) used as a carrier for bone tissue engineering were studied. Following the method described by Urist, allogenic DBM was made. In vitro, DBM and bone marrow stromal cell (BMSC) from rabbits were co-cultured for 3-7 days and subjected to HE staining, and a series of histomorphological observations were performed under phase-contrast microscopy and scanning electron microscopy (SEM). In vivo the mixture of DBM/BMSC co-cultured for 3 days was planted into one side of muscules sacrospinalis of rabbits, and the DBM without BMSC was planted into other side as control. Specimens were collected at postoperative week 1, 2 and 4, and subjected to HE staining, and observed under SEM. The results showed during culture in vitro, the BMSCs adherent to the wall of DBM grew, proliferated and had secretive activity. The in vivo experiment revealed that BMSCs and undifferentiated mesenchymal cells in the perivascular region invaded gradually and proliferated together in DBM/BMSC group, and colony-forming units of chondrocytes were found. Osteoblasts, trabecular bone and medullary cavity appeared. The inflammatory reaction around muscles almost disappeared at the second weeks. In pure DBM group, the similar changes appeared from the surface of the DBM to center, and the volume of total regenerate bones was less than the DBM/BMSC group at the same time. The results indicated that the mixture of DBM and BMSC had good biocompatibility and ectopic induced osteogenic activity.

关键词： bone tissue engineering decalcified bone matrix bone marrow stromal cell biocompatibility

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Effects of Panax notoginseng saponins on hydrogen peroxide-induced apoptosis in cultured rabbit bone marrow stromal cells

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Journal of Pharmaceutical Analysis 2010年第1期22卷 25-29页

作者： Hui Qiang1,2,Guang-Sheng Wang3,Chen Zhang1,Zhi-Bin Shi1,Li-Hong Fan1,Kun-Zheng Wang1 1.Department of Orthopedics,the Second Affiliated Hospital,Medical School of Xi’an Jiaotong University,Xi’an 710004 2.Department of Orthopedics,Shaanxi Province People’s Hospital,Xi’an 710068 3.Department of Orthopedics,Huashan Hospital,Baoji 721000,China. Department of Orthopedics Second Affiliated Hospital Medical School of Xi'an Jiaotong University Xi'an 710004 China Department of Orthopedics Shaanxi Province People's Hospital Xi'an 710068 China Department of Orthopedics Huashan Hospital Baoji 721000 China

Objective To investigate the effects of Panax notoginseng saponins(PNS)on hydrogen peroxide(H2O2)-induced apoptosis in cultured rabbit bone marrow stromal cells(BMSCs).Methods BMSCs from 3-month-old New Zealand rabbits were isolated and cultured by the density gradient centrifugation combined with adherent method.The cultured BMSCs were divided into three groups:normal control,H2O2 treatment(100μmol/L),and PNS pretreatment(0.1g/L).Intracellular reactive oxygen species(ROS)levels as the index of oxidative stress were measured by using 2’7’-dichlorodihydrofluorescein diacetate.Flow cytometry was used to observe the apoptosis of BMSCs by staining with annexinV-FITC/PI.The protein expression of Bax in BMSCs was analyzed by Western blotting.Activity of caspase-3 enzyme was measured by spectrofluorometry.Results Pretreatment with PNS significantly decreased intracellular ROS level induced by H2O2(P<0.01).PNS markedly attenuated H2O2-induced apoptosis rate from 38.68% to 19.24%(P<0.01).PNS reversed H2O2-induced augmentation of Bax expression.Furthermore,PNS markedly reduced the altered in activity of caspase-3 enzyme induced by H2O2(P<0.01).Conclusion PNS has a protective effect on hydrogen peroxide-induced apoptosis in cultured rabbit BMSCs by scavenging ROS and decreasing Bax expression and caspase-3 activity.

关键词： Panax notoginseng saponins reactive oxygen species bone marrow stromal cell apoptosis Bax

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