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Primary Neuronal & Glial culture
The primary cells of the CNS are divided into neurons and glial cells. Neurons are the main signaling cells of the nervous system, and each neuron can form connections with other neuronal cells through synapses.
Glia provides essential functions of neurons (nutrient provision, regulation of ion concentration, mediating immune response…) in various ways, and glial cell types include astrocytes, microglia, and oligodendrocytes.
Primary neurons and glia are used in many fields of neurological research, focusing on cellular mechanisms of the central and peripheral nervous system and neurodegenerative diseases.
Assay Analysis
- Cytotoxic assay: CCK-8, MTT, Live/Dead cell imaging
- Oxidative stress: ROS, Lipid peroxidation
- Mitochondrial Function: TMRM
- Gene expression: RNA sequencing, qPCR
- Protein expression: Western blot, ELISA
Osteoblast / Osteoclast culture
Bone is in a constant state of remodeling, which is important for the maintenance of normal skeletal structure and function. Osteoclasts are responsible for aged bone resorption and osteoblasts are responsible for new bone formation. The resorption and formation is in stable at physiological conditions. However, when the balance is disturbed, bone architecture or function will be abnormal. Bone metabolism diseases, such as osteoporosis or osteopetrosis will occur.
Osteoblast – MC3T3 subclone 4
The MC3T3-E1 cell line established from newborn mouse calvaria is a well-known in vitro osteogenic model system and has been widely used in bone tissue engineering-related research. MC3T3-E1 cells display a sequential development pattern of proliferation and differentiation, resulting in calcified bone tissue similar to in vivo bone formation. The cells differentiate in accordance with the gene activation of osteoblast markers such as alkaline phosphatase (ALP), type I collagen and collagenase-1 (MMP-1), osteonectin (OPN), osteocalcin (OCN).
Assay Analysis
- Cytotoxic assay: CCK-8, MTT
- Osteoblast differentiation: ALP assay
- Gene expression: RNA sequencing, qPCR
- Protein expression: Western blot, ELISA
Osteoclast – Mouse bone marrow cell
Osteoclast are multinucleated bone resorbing cells formed by cytoplasmic fusion of their mononuclear precursors. The Bone marrow-derived macrophages (BMDM) are primary macrophages obtained by in vitro differentiation of bone marrow cells in the presence of macrophage colony-stimulating factor (M-CSF or CSF1) and RANKL.
Assay Analysis
- Cytotoxic assay: CCK-8, MTT
- Osteoclast differentiation: TRAP stain
- Gene expression: RNA sequencing, qPCR
- Protein expression: Western blot
Angiogenesis culture
Angiogenesis, the formation of new vessels from pre-existing endothelium, is an important process in the adult organism because it supports the increasing demands for metabolic supplies (nutrients, various growth factors, and molecular oxygen) at sites of tissue repair or regeneration, during processes such as pregnancy, the female reproductive cycle, wound healing, and revascularization of ischemic tissue. Human umbilical vein endothelial cell (HUVEC) is used to study the function and pathology of endothelial cells. In particular, HUVEC have been used to study VEGF dependent angiogenesis. VEGF promotes the proliferation and migration of endothelial cells, stimulate the vascular permeability.
Assay Analysis
Cell proliferation
- Cell counting
Migration
- Wound healing assay
- Trans well assay (Boyden chamber)
Tube formation:
- Matrigel assay
Gene expression:
- RNA sequencing -> Nanostring
- qPCR
Protein expression: Western blot, ELISA
Other cell culture
Cell line
- Human Peripheral blood mononuclear cell (hPBMC)
- Human Corneal epithelial cell (HCEC)
- SN4741 DA neuronal cell
- Breast cancer cell – MDA-MB-231
- B16F10 cell
Assay Analysis
- Cytotoxic assay: CCK-8, MTT, Live/Dead cell imaging
- Oxidative stress: ROS, Lipid peroxidation
- Mitochondrial Function: TMRM
- Gene expression: RNA sequencing, qPCR
- Protein expression: Western blot, ELISA
- Immunocytochemistry