Thursday, May 6, 2021

Cell migration and invasion-Principles and Methods | Ubigene

Cell migration and invasion

Cell migration, also known as cell crawling, or cell movement, refers to the movement of a cell after receiving a migratory signal or sensing a gradient of certain substances. Cell migration is an alternating process of pseudopodia extension at the cell head, the establishment of new adhesions, and retraction of the cell body tail in a spatiotemporal manner. Cell migration, one of the basic functions of normal cells, is a physiological process of normal growth and development of the organism, and a ubiquitous form of movement of living cells. Cell migration is implicated in processes such as embryonic development, angiogenesis, wound healing, immune responses, inflammatory responses, atherosclerosis, and cancer metastasis.

Cell invasion refers to the ability of cells to migrate from one area to another through the extracellular matrix. Cell invasion is a response of normal and cancer cells to chemical and mechanical stimuli. Before migrating to new areas, the extracellular matrix is degraded by proteases within the cell. Cell invasion often occurs during wound repair, vascularization and inflammatory response as well as abnormal tissue infiltration, tumor cell metastasis, and so on.


Technical Details
Transwell assay is a commonly used to detect the migration ability and invasion ability of cells. Transwell assay, as its name, that is, the compartment with densely packed small holes is put into a well plate (commonly a 24 well plate) . The cell suspension is added to the compartment, which is placed in a 24 well plate with complete medium added. The cells can pass through the holes in the compartment by deformation and run to the outside of the more nutritious compartment and attach to the outside. By staining and counting the cells on the outside of the compartment, it can tell the strength of the migration and invasion ability of the cells.The basic principle of Transwell is to put the small compartment into a culture plate, inside the small compartment called upper compartment, inside the culture plate called lower compartment. The upper and lower culture fluids are separated by a polycarbonate membrane, upper compartment is added by upper culture fluid, lower compartment is added by lower culture fluid. Add the cells to the upper compartment, and because the membrane is permeable, the components in the lower compartment can affect the cells within the upper compartment, so that the effects of the components of culture medium in the lower compartment on cell growth, motility, and so on can be studied.

Tumor migration assay is the study of the migratory ability of tumor cells or, in specific circumstances. Commonly 8.0, 12.0 µ m membrane is used, adding the tumor cells to the upper compartment, and FBS or some specific chemokines are added in the lower compartment. Tumor cells will migrate towards the lower compartment with high nutrient content. Counting the number of cells entering the lower compartment can indicate the migration ability of tumor cells.

Tumor invasion assay is the study of the invasive ability of tumor cells or, in specific circumstances. Unlike the tumor cell migration assay, a layer of Matrigel is additionally added above the polycarbonate membrane to mimic the extracellular matrix. Tumor cells are added to the upper compartment, and if tumor cells want to enter the lower compartment, they must first secrete matrix metalloproteinases (MMPs) to pass through the polycarbonate membrane by digesting the Matrigel. Counting the number of cells that enter the lower compartment can indicate the invasive ability of tumor cells.

Case study

A.Cell migration for human skin melanoma cell line A20578

B.Cell invasion for human skin melanoma cell line A20578


Reference
Yu, Hong et al. “Propofol suppresses proliferation, invasion, and migration of human melanoma cells via regulating microRNA-137 and fibroblast growth factor 9.” Journal of cellular physiology vol. 234,12 (2019): 23279-23288. doi:10.1002/jcp.28896




Service turnaround and deliverables

Service turnaround

5~10 business days, depending on cell growth and experimental design

 

Customer provides

cell lines, drugs, drug treatments and experimental conditions parameter settings;

 

Deliverables

cell lines, drugs, drug treatments and experimental conditions parameter settings;



Other methods to test cell migration and cell invasion



Wednesday, May 5, 2021

Cell Proliferation-Principles and Methods | Ubigene

Cell Proliferation

Cell proliferation is one of the important physiological functions of living cells and is an important life characteristic of organisms. Cell proliferation is the basis of organism growth, development, reproduction as well as heredity.


Technical Details


Normal cells are metabolically vigorous and their mitochondria contain succinate dehydrogenase, which can reduce tetrazolium salts (e.g., MTT, XTT, WST-1, and WST-8, etc.) to purple crystalline substances that deposit around the cell. Then we can use a microplate reader to read the OD value (optical density), thus detecting the status of cell proliferation.

Cell counting kit-8 (also known as CCK-8) is a rapid and highly sensitive assay based on WST-8 (Water Soluble Tetrazolium-8) and widely used for cell proliferation analysis.

The mechanism is: WST-8 [chemical name: 2-(2-Methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium Sodium Salt] is reduced by dehydrogenases in cells under the reaction of the electron carrier 1-methoxy-5-methylphenazinium sulfate dimethyl ester (1-methoxy PMS) to a highly water-soluble yellow formazan product (formazan dye).The number of formazans generated directly correlates to the number of living cells.This property can therefore be exploited to directly carry out cell proliferation assays.The more rapidly the cells proliferate and the more cells proliferate, the darker the color is. For the same cells, there is a linear relationship between the intensity of the color and the number of cells.


Case study


Reference
Deng, Danni et al. “p62 acts as an oncogene and is targeted by miR-124-3p in glioma.” Cancer cell international vol. 19 280. 6 Nov. 2019, doi:10.1186/s12935-019-1004-x


Service turnaround and deliverables

Service turnaround

5~10 business days, depending on cell growth and experimental design

 

Customer provides

cell lines, and experimental conditions parameter settings

 

Deliverables

raw data, analytical results, experimental reports

Other methods to detect cell proliferation



Wednesday, April 28, 2021

Ubigene developed a CRISPR Gene-Editing strategy designer - Red Cotton™

Ubigene developed a CRISPR Gene-Editing strategy designer - Red Cotton™ . Red Cotton™ is based on our actual successfully gene-editing cases for more than 3000 genes, and the related parameters for more than 800 cell lines, combining with the technical advantages of Ubigene-developed CRISPR-U™ for gene KO. Red Cotton™ includes various one-stop tools covering from strategy design to KO validation, and functions such as gRNA design, primer design, vector design, sequence alignment, etc. Ubigene has comprehensively constructed a powerful gene KO system. No matter you are expert or freshly started researchers, Red Cotton™ can make your gene-KO experiment easier!

Red Cotton™ CRISPR Gene Editing Designer

3 ready-to-use KO cell line strategies | 3000 successful cases of gene KO | Parameters of more than 800 cell line

Red Cotton™ CRISPR gene editing system (Red Cotton™ system), developed by Ubigene, is an automatic system with thousands of gene-editing data, bioinformatics and IT technology. You only need to input your target gene and cell line information, and you will get the following information in a minute: 3sets of ready-to-use KO cell line strategies; More than 800 cell line related parameters; 3000 successful cases of gene knockout.

Tools for gene editing

Sequencing data analysis | Primer design | DNA reverse complement | DNA translation | Sequence alignment | Sequence dot-plot | GC content

Red Cotton™ includes various one-stop gene-editing tools, covering from strategy design to KO validation, and functions such as gRNA design, primer design, analysis of sequencing results, etc.

CRISPR-U™

Exclusive innovation, 10 times more efficient in gene-editing.

CRISPR-U™ (based on CRISPR/Cas9 technology), developed by Ubigene, is more efficient than general CRISPR/Cas9 in double-strand breaking, and CRISPR-U™ can greatly improve the efficiency of homologous recombination, easily achieve knockout (KO), point mutation (PM) and knockin (KI) in vitro and in vivo. With CRISPR-U™, Ubigene has successfully modified genes on more than 100 cell lines.

CRISPR/Cas9 recognizes the target sequence with gRNA, and guides Cas9 endonuclease to cut the upstream of PAM, resulting in the double-strand break (DSB) of DNA the target site. To repair the DSB, the cell uses its own DNA repair mechanism to add or delete or replace pieces of DNA sequences via Homology Directed Repair (HDR) or Non-Homologous End Joining (NHEJ).

Red Cotton™ gRNA plasmid bank

Sign-up Red Cotton™ now and over 600 types of gene plasmids are ready for you!

Ubigene has set-up our gRNA plasmid bank, 10000 in-stock plasmids available. Only $80, 3-5 business days!

Ubigene Biosciences

Make genome editing easier

Ubigene Biosciences is co-founded by biological academics and elites from China, the United States, and France. We are located in Guangzhou Science City, which serves as a global center for high technology and innovation. Ubigene Biosciences has 2000㎡ office areas and laboratories, involving genome editing, cell biology technology, and zebrafish research. We provide products and services for plasmids, viruses, cells, and zebrafish. We aim to provide customers with better gene-editing tools for cell or animal research.

Make genome editing easier is the goal of Ubigene. We developed CRISPR-U™ (based on CRISPR/Cas9 technology) which is more efficient than general CRISPR/Cas9 in double-strand breaking, and CRISPR-U™ can greatly improve the efficiency of homologous recombination, easily achieve knockout (KO), point mutation (PM) and knockin (KI) in vitro and in vivo. With CRISPR-U™, Ubigene has successfully edit genes on more than 100 cell lines.

Pragmatism, efficiency, integrity are our business principles. We are committed to providing the best services and products to every customer who chooses us.

Genome Editing Platform
Focusing on the Application of CRISPR-U™ and CRISPR-B™ Gene Editing Technology
1. Provides various types of gene-editing vectors for different species.
2. Provides different virus packaging services, including lentiviruses, adenoviruses and adeno-associated viruses.
3. Provides high-quality services for gene knockout, point mutation, knockin cell lines and microbe.

Cell Biology Platform Cell Biology Platform
Focusing on primary cells
1. Provides over 400 types of primary cells.
2. Provides culture strategies and related products for different cell types.
3. Provides cell biology-related services such as cell isolation, extraction and validation.

Cell gene knockout scheme
Success Cases Cell Solution
Digestive SystemDigestive System
Human Colon Cancer Cell Line HCT 116
Human Colon Cancer Cell Line SW480
Human Colon Cancer Cell Line SW620
Human Colon Cancer Cell Line HCT 116
Human Colon Cancer Cell Line SW480
Human Colon Cancer Cell Line SW620
Human Colon Cancer Cell Line HT-29
Human Colon Cancer Cell Line LoVo
Human colon carcinoma cell line RKO
Human caucasian colon adenocarcinoma cell line COLO 205
Murine colorectal carcinoma cell line CT26.WT
Human colon adenocarcinoma cell line DLD-1
Human colorectal adenocarcinoma cell line NCI-H716
Human colorectal adenocarcinoma cell line Caco-2
Human colon carcinoma cell line T84
Human liver cancer cell line Hep G2
Human hepatocellular carcinoma cell line HuH-7
Human Hepatocellular Carcinoma Cell Line SNU-387
Human hepatobiliary cancer cell line RBE
Human Gastric Cancer Cell Line HGC-27
Human gastric cancer cell line AGS
Human Esophageal Squamous Carcinoma Cell Line KYSE-150
Human Esophageal Squamous Carcinoma Cell Line KYSE-30
Human Colon Cancer Cell Line HCT 116
Human Colon Cancer Cell Line SW480
Human Colon Cancer Cell Line SW620
Human Colon Cancer Cell Line HT-29
Human Colon Cancer Cell Line LoVo
Human colon carcinoma cell line RKO
Human caucasian colon adenocarcinoma cell line COLO 205
Murine colorectal carcinoma cell line CT26.WT
Human colon adenocarcinoma cell line DLD-1
Human colorectal adenocarcinoma cell line NCI-H716
Human colorectal adenocarcinoma cell line Caco-2
Human colon carcinoma cell line T84
Human liver cancer cell line Hep G2
Human hepatocellular carcinoma cell line HuH-7
Human Hepatocellular Carcinoma Cell Line SNU-387
Human hepatobiliary cancer cell line RBE
Human Gastric Cancer Cell Line HGC-27
Human gastric cancer cell line AGS
Human Esophageal Squamous Carcinoma Cell Line KYSE-150
Human Esophageal Squamous Carcinoma Cell Line KYSE-30
Endocrine SystemEndocrine System
Human Breast Cancer Cell Line MDA-MB-231
Human Breast Cancer Cell Line MDA-MB-468
Human Breast Cancer Cell Line MDA-MB-453
Human Breast Cancer Cell Line MDA-MB-436
Rat Breast Cancer Cell Line 4T1
Human Pancreatic Carcinoma Cell Line PANC-1
Human Breast Adenocarcinoma Cell Line SK-BR-3
Human Metastatic Pancreatic Adenocarcinoma Cell Line AsPC-1
Human Pancreatic Carcinoma Cell Line MIA PaCa-2
Human prostate cancer cell line PC-3
Human Prostate Cancer Cell Line VCaP
Mouse Acinar Pancreatic Cell Line 266-6
Human pancreatic cancer cell line BxPC-3
Human Breast Cancer Cell Line MDA-MB-231
Human Breast Cancer Cell Line MDA-MB-468
Human Breast Cancer Cell Line MDA-MB-453
Human Breast Cancer Cell Line MDA-MB-436
Rat Breast Cancer Cell Line 4T1
Human Pancreatic Carcinoma Cell Line PANC-1
Human Breast Adenocarcinoma Cell Line SK-BR-3
Human Metastatic Pancreatic Adenocarcinoma Cell Line AsPC-1
Human Pancreatic Carcinoma Cell Line MIA PaCa-2
Human prostate cancer cell line PC-3
Human Prostate Cancer Cell Line VCaP
Mouse Acinar Pancreatic Cell Line 266-6
Human pancreatic cancer cell line BxPC-3
Respiratory SystemRespiratory System
Human Lung Cancer Cell Line A549
Human Lung Cancer Cell Line Calu-1
Human lung squamous cell carcinoma cell line SK-MES-1
Human Lung Squamous Cell Carcinoma Cell Line NCI-H226
Human Non-small Cell Lung Carcinoma Cell Line NCI-H1299
Human Non-small Cell Lung Carcinoma Cell Line HCC827
Human Bronchial Epithelial Cell Line BEAS-2B
Human Bronchial Epithelial Cell Line 16HBE
Human hypopharyngeal carcinoma cell line FaDu
Human Lung Cancer Cell Line A549
Human Lung Cancer Cell Line Calu-1
Human lung squamous cell carcinoma cell line SK-MES-1
Human Lung Squamous Cell Carcinoma Cell Line NCI-H226
Human Non-small Cell Lung Carcinoma Cell Line NCI-H1299
Human Non-small Cell Lung Carcinoma Cell Line HCC827
Human Bronchial Epithelial Cell Line BEAS-2B
Human Bronchial Epithelial Cell Line 16HBE
Human hypopharyngeal carcinoma cell line FaDu
Blood and Lymphatic SystemBlood and Lymphatic System
Human Monocytic Cell Line THP-1
Rat Basophil Leukemia Cell Line RBL-2H3
Human leukemia cell line HL-60
Human myelogenous leukemia cell line K-562
Human caucasian histiocytic lymphoma cell line U-937
Human Acute Non-B Non-T Lymphocytic Leukemia Cell Line Reh
Human Monocytic Cell Line THP-1
Rat Basophil Leukemia Cell Line RBL-2H3
Human leukemia cell line HL-60
Human myelogenous leukemia cell line K-562
Human caucasian histiocytic lymphoma cell line U-937
Human Acute Non-B Non-T Lymphocytic Leukemia Cell Line Reh
Urinary SystemUrinary System
Human bladder carcinoma cell line TCCSUP
Human bladder carcinoma cell line 5637
Human Bladder Transitional Cell Carcinoma Cell Line T24
Human Embryonic Kidney Cell Line 293T
Human prostate cancer cell line 22RV1
Human bladder carcinoma cell line TCCSUP
Human bladder carcinoma cell line 5637
Human Bladder Transitional Cell Carcinoma Cell Line T24
Human Embryonic Kidney Cell Line 293T
Human prostate cancer cell line 22RV1
Reproductive SystemReproductive System
Human Cervical Carcinoma Cell Line HeLa
Human Cervical Carcinoma Cell Line HeLa 229
Human Cervical Squamous Cell Line SiHa
Human Cervical Carcinoma Cell Line HeLa
Human Cervical Carcinoma Cell Line HeLa 229
Human Cervical Squamous Cell Line SiHa
Chinese Hamster Ovary Cell Line CHO-K1
Human Ovarian Cancer Cell Line SK-OV-3
Human Ovarian Cancer Cell Line OVCAR3
Mouse Pituitary Cell Line Lbetat2
Mouse Testicular Stromal Cell Line TM3
Circulatory SystemCirculatory System
Mouse Myoblast Cell Line C2C12
Brain and Nervous SystemBrain and Nervous System
Rat Glioblastoma Cell Line C6
Mouse Glioblastoma Cell Line GL261
Human glioblastoma cell line U-87 MG
Mouse Anterior Parietal Bone Cell Line MC3T3-E1 Subclone 14
Mouse Hippocampal Neuron Cell Line HT22
Human Neuroblastoma Cell Line SK-N-SH
Mouse neuroblastoma cell line Neuro-2a
Human Glioblastoma Cell Line U251
Rat Glioblastoma Cell Line C6
Mouse Glioblastoma Cell Line GL261
Human glioblastoma cell line U-87 MG
Mouse Anterior Parietal Bone Cell Line MC3T3-E1 Subclone 14
Mouse Hippocampal Neuron Cell Line HT22
Skeleton, Articulus, Soft Tissue, Derma SystemSkeleton, Articulus, Soft Tissue, Derma System
Human Osteosarcoma Cell Line MG-63
Human bone osteosarcoma epithelial cell line U-2 OS
Human fibrosarcoma cell line HT-1080
Human malignant melanoma cell line A-375
Human Melanoma Cell Line M14
Murine melanoma cell line B16-F10
Human Osteosarcoma Cell Line MG-63
Human bone osteosarcoma epithelial cell line U-2 OS
Human fibrosarcoma cell line HT-1080
Human malignant melanoma cell line A-375
Human Melanoma Cell Line M14
Murine melanoma cell line B16-F10
Ocular, Otolaryngologic and Oral SystemOcular, Otolaryngologic and Oral System
Human Nasopharyngeal Carcinoma Cell Line CNE2Z
Human Nasopharyngeal Carcinoma Cell Line CNE2Z
Microbial gene knockout design strategy

Preferential promotion services
  • Gene-KO cell line
  • KI and Point Mutation Cell Line
  • Red Cotton™ gRNA plasmid bank
  • Stable cell line .Virus packaging

Cell migration and invasion-Principles and Methods | Ubigene

Cell migration and invasion Cell migration, also known as cell crawling, or cell movement, refers to the movement of a cell after receiving ...