**4.2 In vivo analyses**

*Update on Mesenchymal and Induced Pluripotent Stem Cells*

networks in ischemic regions [126, 127].

sive cardiac regeneration.

**4. Angiogenesis assays**

**4.1 In vitro analyses**

*4.1.2 Migration assays*

*4.1.4 Aortic ring assay*

periphery [132].

*4.1.1 Proliferation and survival assays*

MCP-1) TGF-β, neurotrophic factors (brain-derived neurotrophic factor (BDNF)),

Exosomes from MSCs exposed to hypoxia and FBS-free condition enhanced neovascularization in the injured heart [92, 122–124]. In a preclinical study, intramyocardial transplantation of exosome secreted from MSCs significantly improves blood flow rate and reduced infarct zone in the rat model [125]. Approximately, the entire small and large animal model of CVD preclinical investigations along with high-quality phase 0, I, II, and III clinical trials and meta-analysis studies vigorously confirmed that MSC therapy has the effective effects in developing angiogenic

Ongoing researches on preconditioning and genetic manipulations of MSCs are needed to enhance angiocrine capacity governed by growth factors, microvesicles, microRNAs, long noncoding RNAs (lncRNAs), etc. [128, 129]. Finding the route of cell delivery, the optimum dose, the excellent cell source, and transplantation time are factors that still require to be addressed so as to achieve the aim of comprehen-

Both in vitro and in vivo angiogenesis assays are commonly used to investigate

Monitoring the proliferation of ECs is needed to develop microvascular units. Different survival and proliferation assays based on DNA synthesis or metabolic status are applicable. These assays could also predetermine the anti-angiogenic

This method shows the migration in response to diverse factors, ability to digest basal membrane, and healing capacity of MSCs which is done by various assays as follows: Boyden chamber assay, Transwell® inserts, agarose assay, wound-healing

This system is done in the 2D and 3D milieu and able to monitor alignment and juxtacrine connection of cells after plating on a specific substrate such as Matrigel, Fibrin, etc. Plated cells acquire phenotype to form capillary-like structures and lumen which are applicable to in vivo condition and evaluated in terms of tube area

In this assay, the aorta from mouse or rats was removed and placed on collagen or fibrin matrix in serum-free condition. The angiogenic potential is determined by EC sprouting, polarized cells, and outgrowth appearance to the

pro- and anti-angiogenic potential of stem cells and different cell types.

property of a specific compound in the context of tumor biology.

assay, Teflon fence assay, phagokinetic track, etc. [130].

*4.1.3 Tube formation (tubulogenesis) assay*

and number per microscopic field [130, 131].

nitric oxide (NO), and improved cardiac restoration after injury [121].

**110**
