Expansion and characterization of mesenchymal stem cells and other anchorage-dependent cell types in fluoropolymer bags treated for adherent cell culture

Of all the significant researches that have taken place in isolation, culture and characterization of mesenchymal stem cells (MSCs), the field of kidney-derived mesenchymal stem cells (KMSCs) in Tibetan mastiff is still a blank. Therefore, the purpose of this study is to isolate, culture and characterize the Tibetan mastiff KMSCs. The KMSCs were successfully isolated from one-day year old Tibetan mastiff kidney, cultured for 16 passages and distinguished by two methods: immunofluorescence staining and RT-PCR. The Tibetan mastiff KMSCs expressed specific surface marker genes (VIM, CD44, FN1, CD90, CD109, CD73, FN1) and kidney marker gene PAX2. The proliferation ability of Tibetan mastiff KMSCs was measured through cell count and clonality. Furthermore, cells differentiated into different cell types (hepatocellular cells, osteogenic cells, adipogenic cells and chondrogenic cells) under special induced medium, and the marker genes of induced cells were identified with Immunofluorescence staining and RT-PCR. All of these results indicated that the Tibetan mastiff KMSCs were obtained successfully, which possessed certain characteristics of multipotent stem cells. Therefore, MSCs in Tibetan mastiff kidney hold potential for clinical applications for regenerative therapy and their further studies are waiting to be required to investigate their functions.

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Laminin-adherent versus suspension-non-adherent cell culture conditions for the isolation of cancer stem cells in the DAOY medulloblastoma cell line

Tumor Biology ◽

10.1007/s13277-016-5119-6 ◽

2016 ◽

Vol 37 (9) ◽

pp. 12359-12370 ◽

Cited By ~ 4

Author(s):

Javier de la Rosa ◽

Ander Sáenz Antoñanzas ◽

Mehdi H. Shahi ◽

Bárbara Meléndez ◽

Juan A. Rey ◽

...

Keyword(s):

Stem Cells ◽

Cell Culture ◽

Cancer Stem Cells ◽

Cell Line ◽

Culture Conditions ◽

Adherent Cell ◽

Medulloblastoma Cell ◽

Medulloblastoma Cell Line ◽

Adherent Cell Culture ◽

Cell Culture Conditions

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Establishment and biological characterization of a dermal mesenchymal stem cells line from bovine

Bioscience Reports ◽

10.1042/bsr20130095 ◽

2014 ◽

Vol 34 (2) ◽

Cited By ~ 7

Author(s):

Tingting Sun ◽

Chao Yu ◽

Yuhua Gao ◽

Chenqiong Zhao ◽

Jinlian Hua ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Cell Types ◽

Neural Cells ◽

Biological Characterization ◽

Multipotent Stem Cells ◽

Reverse Transcription Pcr ◽

Lineage Differentiation

The DMSCs (dermal mesenchymal stem cells) are multipotent stem cells, which can differentiate in vitro into many cell types. Much work has been done on DMSCs from humans, mice, rabbits and other mammals, but the related literature has not been published about these cells in cattle. In this study, we isolated and established the DMSC lines from cattle, thereby initiating further research on these cells, such as growth kinetics, detection of special surface antigen and RT–PCR (reverse transcription–PCR) assays to identify the biological characterization of the cell line. Furthermore, the DMSCs are induced to differentiate into adipocytes, osteoblasts and neural cells in vitro. Our results suggest that DMSCs isolated from cattle possess similar biological characteristics with those from other species. Their multi-lineage differentiation capabilities herald a probable application model in tissue engineering and induced pluripotent stem cells.

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Automated and Online Characterization of Adherent Cell Culture Growth in a Microfabricated Bioreactor

Journal of Laboratory Automation ◽

10.1177/2211068214529288 ◽

2014 ◽

Vol 19 (5) ◽

pp. 437-443 ◽

Cited By ~ 15

Author(s):

Nicolas Jaccard ◽

Rhys J. Macown ◽

Alexandre Super ◽

Lewis D. Griffin ◽

Farlan S. Veraitch ◽

...

Keyword(s):

Cell Culture ◽

Adherent Cell ◽

Adherent Cell Culture ◽

Culture Growth ◽

Cell Culture Growth

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Spheroid Culture of Mesenchymal Stem Cells

Stem Cells International ◽

10.1155/2016/9176357 ◽

2016 ◽

Vol 2016 ◽

pp. 1-11 ◽

Cited By ~ 138

Author(s):

Zoe Cesarz ◽

Kenichi Tamama

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Cell Biology ◽

Molecular Mechanisms ◽

Replicative Senescence ◽

Marker Genes ◽

Adherent Cell ◽

Pluripotency Marker ◽

Adherent Cell Culture ◽

Microarray Analyses

Compared with traditional 2D adherent cell culture, 3D spheroidal cell aggregates, or spheroids, are regarded as more physiological, and this technique has been exploited in the field of oncology, stem cell biology, and tissue engineering. Mesenchymal stem cells (MSCs) cultured in spheroids have enhanced anti-inflammatory, angiogenic, and tissue reparative/regenerative effects with improved cell survival after transplantation. Cytoskeletal reorganization and drastic changes in cell morphology in MSC spheroids indicate a major difference in mechanophysical properties compared with 2D culture. Enhanced multidifferentiation potential, upregulated expression of pluripotency marker genes, and delayed replicative senescence indicate enhanced stemness in MSC spheroids. Furthermore, spheroid formation causes drastic changes in the gene expression profile of MSC in microarray analyses. In spite of these significant changes, underlying molecular mechanisms and signaling pathways triggering and sustaining these changes are largely unknown.

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Using an adherent cell culture of the mouse subependymal zone to study the behavior of adult neural stem cells on a single-cell level

Nature Protocols ◽

10.1038/nprot.2011.404 ◽

2011 ◽

Vol 6 (12) ◽

pp. 1847-1859 ◽

Cited By ~ 25

Author(s):

Felipe Ortega ◽

Marcos R Costa ◽

Tatiana Simon-Ebert ◽

Timm Schroeder ◽

Magdalena Götz ◽

...

Keyword(s):

Stem Cells ◽

Cell Culture ◽

Neural Stem Cells ◽

Single Cell ◽

Adherent Cell ◽

Single Cell Level ◽

Cell Level ◽

Adult Neural Stem Cells ◽

Subependymal Zone ◽

Adherent Cell Culture

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Identification of Novel FNIN2 and FNIN3 Fibronectin-Derived Peptides That Promote Cell Adhesion, Proliferation and Differentiation in Primary Cells and Stem Cells

International Journal of Molecular Sciences ◽

10.3390/ijms22063042 ◽

2021 ◽

Vol 22 (6) ◽

pp. 3042

Author(s):

Eun Ju Lee ◽

Khurshid Ahmad ◽

Shiva Pathak ◽

SunJu Lee ◽

Mohammad Hassan Baig ◽

...

Keyword(s):

Stem Cells ◽

Cell Culture ◽

Cell Adhesion ◽

3D Culture ◽

Human Mesenchymal Stem Cells ◽

Cell Types ◽

Primary Cells ◽

Cell Culture Techniques ◽

Culture Techniques ◽

Promote Cell Adhesion

In recent years, a major rise in the demand for biotherapeutic drugs has centered on enhancing the quality and efficacy of cell culture and developing new cell culture techniques. Here, we report fibronectin (FN) derived, novel peptides fibronectin-based intergrin binding peptide (FNIN)2 (18-mer) and FNIN3 (20-mer) which promote cell adhesion proliferation, and the differentiation of primary cells and stem cells. FNIN2 and 3 were designed based on the in silico interaction studies between FN and its receptors (integrin α5β1, αvβ3, and αIIbβ3). Analysis of the proliferation of seventeen-cell types showed that the effects of FNINs depend on their concentration and the existence of expressed integrins. Significant rhodamine-labeled FNIN2 fluorescence on the membranes of HeLa, HepG2, A498, and Du145 cells confirmed physical binding. Double coating with FNIN2 or 3 after polymerized dopamine (pDa) or polymerized tannic acid (pTA) precoating increased HBEpIC cell proliferation by 30–40 percent, suggesting FNINs potently affect primary cells. Furthermore, the proliferation of C2C12 myoblasts and human mesenchymal stem cells (MSCs) treated with FNINs was significantly increased in 2D/3D culture. FNINs also promoted MSC differentiation into osteoblasts. The results of this study offer a new approach to the production of core materials (e.g., cell culture medium components, scaffolds) for cell culture.

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