siRNA / miRNA gene silencing Human PEL

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.

RNA-Seq is a method to sequence RNA by applying Next Generation Sequencing (NGS). The quality of RNA is critical for the success of RNA-Seq. The integrity of RNA is measured by the RNA integrity number (RIN). RIN is computed from RNA electrophoresis and electropherogram profiles (the peak area of the 28S rRNA should be approximately twice the peak area of the 18S rRNA). If you get the RIN value lower than 7, the possibility of getting the low quality of RNA-seq data is high. To get a high quality RNA, it is better to work with fresh samples or snap-freeze the tissues in liquid nitrogen as quickly as possible and store them at -80°C until further use. Make sure designated areas and all your filter tips, microfuge tubes, plastic, and glassware are RNase-free.

Get tips on using ScriptSeq Complete Kit (Human/Mouse/Rat) to perform RNA sequencing Mouse - Bone marrow-derived macrophages (BMDMs)

Get tips on using Polyclonal Rabbit Anti-Human Myeloperoxidase (Dako Omnis) to perform Immunohistochemistry Mouse - MPO

Get tips on using Purified Mouse Anti-Human MLH1 Clone G168-728 (RUO) to perform Immunohistochemistry Human - MLH1

Get tips on using Monoclonal Mouse Anti-Human Androgen Receptor (Concentrate) Clone AR441 to perform Immunohistochemistry Human - AR

Get tips on using Monoclonal Mouse Anti-Human Cytokeratin 20 (Concentrate) Clone Ks20.8 to perform Immunohistochemistry Human - CK20

Get tips on using Human Sonic Hedgehog/Shh N-Terminus Quantikine ELISA Kit to perform ELISA Human - ShhN

Get tips on using Human Retinol binding protein 4 ELISA Kit (RBP4) (ab108897) to perform ELISA Human - RBP4

Get tips on using Human NRG1-beta 1/HRG1-beta 1 DuoSet ELISA to perform ELISA Human - NRG1