siRNA / RNAi /miRNA transfection Human Cells A549

Stem cells have the unique ability to self-renew or differentiate themselves into various cell types in response to appropriate signals. These cells are especially important for tissue repair, regeneration, replacement, or in the case of hematopoietic stem cells (HSCs) to differentiate into various myeloid populations. Appropriate signals refer to the growth factor supplements or cytokines that mediate differentiation of various stem cells into the required differentiated form. For instance, HSCs can be differentiated into dendritic cells (with IL-4 and GM-CSF), macrophages (with m-CSF) and MDSCs (with IL-6 and GM-CSF). Human pluripotent stem cells (hPSCs) and induced pluripotent stem cells (iPSCs) can be first cultured in neural differentiation media (GSK3𝛃-i, TGF𝛃-i, AMPK-i, hLIF) to form neural rosettes, which can be differentiated into neural or glial progenitors (finally differentiated into oligodendrocytes). Neural progenitors can be finally differentiated into glutaminergic (dibytyryl cAMP, ascorbic acid) and dopaminergic (SHH, FGF-8, BDNF, GDNF, TGF-𝛃3) neurons. Thus, it is important to first identify the self-renewing cell line: its source and its final differentiation state, followed by the supplements and cytokines required for the differentiation, and final use. Timelines are another thing that is considered. For instance, it takes 7-10 days to form neural rosettes from iPSCs and 3 days to differentiate neural progenitors to neurons. Finally, the stability for stem cell culture media varies. It is advised to make fresh media every time when differentiating HSCs to myeloid populations, whereas neural differentiation media may remain stable for two weeks when stored in dark between 2-8C.

Get tips on using Flot1 Rat siRNA Oligo Duplex (Locus ID 64665) to perform siRNA / miRNA gene silencing Rat - NRCM Flot1

Get tips on using Flot2 Rat siRNA Oligo Duplex (Locus ID 83764) to perform siRNA / miRNA gene silencing Rat - NRCM Flot2

Get tips on using Dynabeads™ Untouched™ Human T Cells Kit to perform Cell Isolation Human T cells

Wound healing assay can be challenging due to inconsistencies and variations while making a wound on the confluent cell monolayer, consequently leads to wounds of varying sizes and widths. Moreover, this assay causes damage to the cells that are at the edge of the wound, which can prevent cell migration into the wound site and healing. The best solution is to use the standard wound healing assay kits using either combs or inserts to make a defined wound field or gap and prevent the well-to-well variation in these assays.

Get tips on using ON-TARGETplus Rat Prkaa1 (65248) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - UMR‐106 Ampkα1/Prkaa1

Get tips on using ON-TARGETplus Rat Prkaa2 (78975) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - UMR‐106 Ampkα2/Prkaa2

Get tips on using ON-TARGETplus Rat Arhgef6 (363509) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - MTLn3 α-Pix/Arhgef6

Get tips on using ON-TARGETplus Rat Arhgef7 (114559) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - MTLn3 β-Pix/Arhgef7

Get tips on using ON-TARGETplus Rat Itgb1 (24511) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - MTLn3 β1 integrin/Itgb1