Essential role of ATP6AP2 enrichment in caveolae/lipid raft microdomains for the induction of neuronal differentiation of stem cells

Makdissy, Nehman; Haddad, Katia; AlBacha, Jeanne D’arc; Chaker, Diana; Ismail, Bassel; Azar, Albert; Oreibi, Ghada; Ayoub, David; Achkar, Ibrahim; Quilliot, Didier; Fajloun, Ziad

doi:10.1186/s13287-018-0862-9

Stem Cell Research & Therapy

Table 1 Immunophenotyping, apoptotic index and distribution of neurogenic markers on unpurified mixed population

From: Essential role of ATP6AP2 enrichment in caveolae/lipid raft microdomains for the induction of neuronal differentiation of stem cells

	MSCs Day < 0	NSPs Day 30	NLCs Day 37
Differentiation of hADSCs in healthy subjects
CD34⁽⁺⁾	1.26 ± 2.01	0.18 ± 1.64	0.51 ± 2.60
CD45⁽⁺⁾	1.28 ± 1.17	0.71 ± 0.92	0.07 ± 1.34
CD73⁽⁺⁾	90.12 ± 5.52	24.51 ± 10.34 ^*	6.77 ± 3.68 ^**
CD90⁽⁺⁾	95.59 ± 6.80	4.07 ± 3.76 ^**	1.94 ± 2.1 ^***
CD105⁽⁺⁾	84.33 ± 5.33	18.68 ± 4.51 ^*	1.26 ± 1.90 ^***
Annexin V⁽⁻⁾/PI⁽⁻⁾/7AAD⁽⁻⁾ (viable cells)	98.97 ± 0.71	96.20 ± 1.25	96.85 ± 0.83
Annexin V⁽⁺⁾/PI⁽⁻⁾/7AAD⁽⁻⁾ (early-apoptotic cells)	0.76 ± 0.90	2.11 ± 1.01	2.03 ± 1.04
Annexin V⁽⁺⁾/PI⁽⁺⁾/7AAD⁽⁺⁾ (late apoptotic, necrotic cells)	0.27 ± 0.71	1.69 ± 0.99	1.12 ± 1.10
NeuN	1.62 ± 1.54	58.14 ± 3.32 ^**	78.12 ± 6.28 ^**
TUJ1	4.49 ± 1.62	60.48 ± 5.25 ^**	89.09 ± 8.67 ^**
O4	1.28 ± 2.40	28.16 ± 4.11^*	26.33 ± 3.29 ^*
GFAP	24.14 ± 2.58	4.26 ± 1.84 ^**	1.20 ± 1.24 ^**
Nestin	54.01 ± 8.41	20.15 ± 2.64 ^*	0.81 ± 2.62 ^**
Differentiation of hADSCs in ND subjects
CD34⁽⁺⁾	1.82 ± 1.72	0.08 ± 1.21	0.71 ± 1.05
CD45⁽⁺⁾	3.69 ± 1.88	0.90 ± 1.14	0.11 ± 1.92
CD73⁽⁺⁾	79.49 ± 3.42	4.06 ± 2.66 ^**	2.90 ± 2.03 ^***
CD90⁽⁺⁾	93.51 ± 3.01	0.73 ± 1.40 ^***	0.98 ± 1.74 ^***
CD105⁽⁺⁾	86.28 ± 6.76	13.53 ± 5.18 ^**	5.04 ± 7.18 ^**
Annexin V⁽⁻⁾/PI⁽⁻⁾/7AAD⁽⁻⁾ (viable cells)	97.12 ± 1.13	93.10 ± 2.44	98.33 ± 2.91
Annexin V⁽⁺⁾/PI⁽⁻⁾/7AAD⁽⁻⁾ (early apoptotic cells)	1.96 ± 0.76	4.71 ± 1.82	0.02 ± 2.16
Annexin V⁽⁺⁾/PI⁽⁺⁾/7AAD⁽⁺⁾ (late apoptotic, necrotic cells)	0.92 ± 1.33	2.19 ± 1.37	1.65 ± 0.24
NeuN	0.37 ± 1.09	45.20 ± 2.07 ^**	50.31 ± 2.53 ^{**, §}
TUJ1	1.94 ± 1.13	40.11 ± 4.17 ^{**, §}	52.65 ± 6.24 ^{**, §§}
O4	4.13 ± 5.10	38.14 ± 5.23^**	18.04 ± 1.88 ^{*, §}
GFAP	34.05 ± 3.67 ^§	4.19 ± 2.12 ^**	2.43 ± 4.16 ^**
Nestin	73.56 ± 7.82 ^§	16.20 ± 3.75 ^**	1.05 ± 2.35 ^**

Cells were labeled with fluorescence-coupled antibodies against CD34, CD45, CD73, CD90, CD105, Annexin V, 7AAD, propidium iodide solution (PI) and neural markers and analyzed using a MACSQuant flow analyzer as indicated in Methods. Results presented as mean ± SEM and presented as the percentage of cell surface marker per cell type (% of total cells) of all subjects each performed in duplicate in each group. Cells collected at day 0 before the induction of the differentiation (MSCs) and days 30 and 37 of differentiation (NSPs and NLCs, respectively)
hADSC human adipose-derived mesenchymal stem cell, MSC mesenchymal stem cell, ND neurodegenerative disease, NLC neural-like cell, NSP neurosphere-like structure
^*P < 0.05, ^**P < 0.01 and ^***P < 0.005: NSPs/NLCs vs MSCs
^§P < 0.05, ^§§P < 0.01: ND vs healthy

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ISSN: 1757-6512

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