Hi to everyone.I want to know some microRNAs that enhance osteogenesis and chondrogenesis?

Can anyone share their experience with C3H10T1/2 cells for chondrogenic and/or osteogenic induction, please? In particular, the efficacy/efficiency of differentiation and the timing.

I am currently osteochondral differentiation of MSCs but would like to use a cell line instead of primary cells (i.e. from the bone marrow). I believe that C3H10T1/2s are embryo-derived so I am a little in doubt of their efficacy for differentiation into chondrocytes and osteblasts. For inducing differentiation, I use the standard chondrogenic and osteogenic medium (including ascorbic acid, DEX, b-glycerophosphate, etc). For my experiments, I only culture the samples for 1 or 2 weeks max. Is this too short to see the differentiation??

Thanks!

I want to make a skeletal staining for E15.5 and E18.5 mouse embryos. Some publications refer to a method described by Kimmel and Trammell (Stain Technol. 1981). Does anyone have experience using this method, or can give any clues for an adequately fast (2-5 days) procedure?

I´m working on a project with 3-lineage differentiation of equine MSCs from adipose tissue and peripheral blood. I differentiate them towards catilage, bone and adipose tissue. In the following you can see the media and supplements I use.

My question is what exatly is the effect of dexamethasone to the cells, since the supplements is used for all 3 lineages it cannot be for the differentiation towards a specific lineage. Also what is the excat effect of asorbic acid (it generally seems to be used in a much higher density in medias towards osteogenesis than towards chondrogenesis), beta-glycerophosphate, TGFbeta, insulin and IBMX, indometahcin.

Or maybe you know a good place to find information about the exact effect of these supplements. Im very new i this field, so sorry if the question is a little stupid.  

Monolayer, DMEM-LG, FCS, 1% pen/strep,50 µg/ml gentamycin

0,1 µM dexamethasone

50 µM L-ascorbicacid-2-phosphate

10 mM β-glycerophosphate

Chondrogenesis 

3D pellets, DMEM-HG, 10% FCS, 1% pen/strep,50 µg/ml gentamycin

0,1 µM dexamethasone

1 µM L-ascorbicacid-2-phosphate

10 ng/ml TGFβ3

Adipogenesis 

Monolayer, DMEM-LG, 15% Rabbitserum, 1% pen/strep, 50 µg/ml gentamycin

1 µM dexametason

0,5 mM IBMX

10 µg/ml rh insulin

0,2 mM indomethacin

all are stored in a incubator with 37 degree C, and 5 % CO2. 

I just have started my project. My project focuses on the BMP-2 and VEGF role in osteogenesis and vascularization. I am doing survey looking for enzymes that cleaving proBMP-2 and proVEGF. I am interesting in finding the sequences that make BMP-2 and VEGF inactive initially and active ultimately by enzymes secreted from osteoprogenitor cells. If you could help me in that I will appreciate that

Thanks in advance.

In comparison to differentiation of MSCs towards adipogenis and osteogenesis,  most literature shows to use  media with a high content of glucose for chondrogenesis. Why is that? 

Some believe it is due to promote the effect of TGFbeta but could it also be due to  different demands on the cells when grown as a pellet? 

we take a XRay in a conservative treatement u see callus and decide its united

when u do plate osteisynthesis when there is no callus how to decide if there is a gap due to commuintion and whther the pixel densith in a x-ray can be used to decide there is caluus formation or bone formation or not?

I read papers about in vitro transcribed RNA. What is the difference between in vitro transcribed RNA and commercially purchased RNA?

Thanks

Cementoid, new bone and new Cementum is present histologically .