Table 2.

Summarizes the studies that apply hypoxia for studying changes in clonogenic potential of MSCs. These studies have reported that the clonogenic potential of MSCs is enhanced under hypoxic culture conditions

Reference	Material	Conditions and device used	Cell Source	Medium/Growth Factors Used	Reported Result
Boyette et al.[ 5 ]	6‐well or 24‐well plates	Hypoxic culture (5% O2) for 21 days in closed incubators;	Human bone marrow derived MSC (hbMSC) Cell density 1 × 104 cells cm−2	Basal medium (High glucose DMEM supplemented with 10% FBS and antibiotics) for expansion	Increase in clonogenicity due to increased cell proliferation, increased secretion of VEGF, and increased matrix turnover
Antebi et al.[ 32 ]	Tissue culture flasks	Hypoxic culture (1% O2) for long‐term (10 days) and short‐term (48 h). Or Short‐term (48 h) 2% and 5% O2 in hypoxia station (HypOxystation H35, HypOxygen)	hbMSC and procine bMSC Cell density 3 × 105 cells cm−2	Basal medium (α‐MEM supplemented with 15% FBS. 2 × 10−3 m L‐glutamine, and antibiotics)	Slower proliferation and lower yields of MSCs under long term exposure to hypoxia. Short term hypoxic culture led to significantly faster proliferation. Increased clonogenic potential due to increased expression of VEGF and decreased expression of apoptotic genes BCL‐2 and CASP3 in both short and long term hypoxic exposures.
Li et al.[ 36 ]	Tissue culture flasks	Hypoxic culture (2.5% O2) in a hypoxia gas chamber for 5 days	Mouse bone marrow derived MSCs (mbMSC)	Basal medium (DMEM/F12 supplemented with 10% FBS and antibiotics)	mbMSCs exposed to hypoxia had higher cell viability and proliferation potential Hypoxic mbMSCs showed increased clonogenic potential alongwith increased cell proliferation
Krinner et al.[ 28 ]	96‐well plates for clonal expansion assay	Hypoxic culture (5% O2) for 14 days in tri‐gas incubator (Thermo Fisher Scientific)	Sheep bone marrow derived MSC (sbMSC) Cell density 1 cell/96‐well for clonal expansion	Basal medium (High‐glucose DMEM supplemented with 10% FCS and antibiotics)	Alongwith proliferation, hypoxia increased clonogenicity and colony forming ability of ovine MSCs.
Hu et al.[ 29 ]	6‐well plates for clonal expansion assay	Hypoxic culture (5% O2 and 10% O2) for 14 days.	mbMSC Cell density 1 × 104 cells/96‐well for clonal expansion	Basal medium (DMEM with 1500 mg L−1 D‐glucose, 20% FBS, 1% glutamine and antibiotics)	Mice bMSCs showed enhanced clonogenicity and colony forming ability at 5% O2 compared to 10% O2 and normoxia.
Adesida et al.[ 37 ]	T150 tissue culture flask for clonal expansion	Hypoxic culture (3% O2) for 14 days	hbMSC Cell density 100 000 cells cm−2 for expansion	Basal medium (α‐MEM supplemented with 10% FBS, 4‐(2‐hydroxyethyl)‐1‐piperazineethanesulfonic acid (HEPES), sodium pyruvate, 5 ng mL−1 basic fibroblast growth factor (bFGF) and antibiotics)	Significantly higher number of hbMSC colonies under hypoxia as compared to normoxia.