<i>In vitro</i> maturation medium supplementation: utilization of repaglinide, L-carnitine, and mesenchymal stem cell-conditioned medium to improve developmental competence of oocytes derived from endometriosis mouse models

Kalehoei, E.; Moradi, M.; Azadbakht, M.; Zhaleh, H.; Parvini, M.; Cheraghbaeigi, S.; Saghari, S.

doi:10.1590/1414-431X2022e11948

Acessibilidade / Reportar erro

Brasil

Brazilian Journal of Medical and Biological Research

Español English

Brasil

Español English

sumário « anterior atual seguinte »

Sumário

Research Article • Braz. J. Med. Biol. Res. 55 • 2022 • https://doi.org/10.1590/1414-431X2022e11948 copy

In vitro maturation medium supplementation: utilization of repaglinide, L-carnitine, and mesenchymal stem cell-conditioned medium to improve developmental competence of oocytes derived from endometriosis mouse models

Authorship SCIMAGO INSTITUTIONS RANKINGS

Endometriosis (EMS) is one of the most prevalent causes for female infertility. Herein, we investigated the effect of the repaglinide (RG), L-carnitine (LC), and bone marrow mesenchymal stem cell-conditioned medium (BMSC-CM) supplementation during in vitro maturation (IVM) on the quality, maturation, and fertilization rates, as well as embryonic quality and development of oocytes derived from normal and EMS mouse model. Immature oocytes were collected from two groups of normal and EMS-induced female NMRI mice at 6-8 weeks of age. Oocytes were cultured in IVM medium unsupplemented (control group), or supplemented with 1 M RG, 0.3 and 0.6 mg/mL LC, and 25 and 50% BMSC-CM. After 24 h of oocyte incubation, IVM rate and antioxidant status were assessed. Subsequently, the rates of fertilization, cleavage, blastulation, and embryonic development were assessed. Our results demonstrated that supplementation of IVM medium with LC and BMSC-CM, especially 50% BMSC-CM, significantly enhanced IVM and fertilization rates, and markedly improved blastocyst development and total blastocyst cell numbers in EMS-induced mice compared to the control group (53.28±0.24 vs 18.09±0.10%). Additionally, LC and BMSC-CM were able to significantly modulate EMS-induced nitro-oxidative stress by boosting total antioxidant capacity (TAC) and mitigating nitric oxide (NO) levels. Collectively, LC and BMSC-CM supplementation improved oocyte quality and IVM rates, pre-implantation developmental competence of oocytes after in vitro fertilization, and enhanced total blastocyst cell numbers probably by attenuating nitro-oxidative stress and accelerating nuclear maturation of oocytes. These outcomes may provide novel approaches to refining the IVM conditions that can advance the efficiency of assisted reproductive technologies in infertile couples.

Endometriosis; Oocyte quality; Maturation culture medium; Bone marrow mesenchymal stem cell; Oxidative stress; Female infertility

Groups	N	GV (%)	GVBD (%)	MII (%)	DEG (%)
Normal
Co	112	16.48±1.07^b,c,d,e,f	21.03±0.85^c,d,e,f	55.16±1.03^b,c,d,e,f	7.25±0.76
RG	108	10.46±1.01^a,e,f	20.87±1.02^c,d,e,f	62.91±1.11^a,c,d,e,f	5.69±0.88
LC-0.3	110	10.12±0.77^a,e,f	16.73±1.00^{a,b c,d,e,f}	68.35±0.84^a,b,d,e,f	4.71±0.55
LC-0.6	114	8.44±0.86^a,e,f	15.19±0.77^a,b,c,f	71.14±0.69^a,b,c,e,f	5.16±0.90
CM-25%	110	5.37±0.91^a,b,c,d	14.52±1.06^a,b,c,f	75.22±1.01^a,b,c,d,f	4.82±1.03
CM-50%	112	5.24±1.02^a,b,c,d	10.23±0.65^a,b,c,d,e	81.59±0.93^a,b,c,d,e	4.14±0.88
Endometriosis
Co	105	18.43±0.089^b,c,d,e,f	35.53±0.015^e,f	39.13±0.022^c,d,e,f	5.40±0.011
RG	102	12.72±0.086^a,f	38.77±0.008^e,f	38.91±0.012^c,d,e,f	9.22±0.016^c,d
LC-0.3	107	12.52±0.066^a,f	35.35±0.007^e,f	50.45±0.025^a,b,e,f	1.96±0.013^b,f
LC-0.6	105	8.31±0.046^a	35.21±0.004^e,f	54.14±0.022^a,b,e,f	3.34±0.023^b
CM-25%	106	9.28±0.044^a	16.25±0.011^a,b,c,d	69.13±0.014^a,b,c,d	4.63±0.028
CM-50%	104	7.14±0.018^a,b,c	8.83±0.018^a,b,c,d,e	76.86±0.009^a,b,c,d,e	7.11±0.031^c

Groups	NO (nmol/mg)	TAC (nmol/mg)
Normal
Co	48.09±1.89^d,e,f	0.97±0.093^c,d,e,f
RG	50.00±1.31^c,d,e,f	0.90±0.046^c,d,e,f
LC-0.3	42.72±1.97^b	1.22±0.079^a,b,d
LC-0.6	39.58±1.09^a,b	1.48±0.058^a,b,c
CM-25%	39.92±1.49^a,b	1.30±0.056^a,b
CM-50%	38.76±1.51^a,b	1.29±0.070^a,b
Endometriosis
Co	85.42±1.12^c,d,e,f	0.52±0.022^c,d,e,f
RG	88.34±1.36^c,d,e,f	0.48±0.036^c,d,e,f
LC-0.3	70.21±1.04^a,b	0.74±0.046^a,b
LC-0.6	62.38±1.2^a,b,e	0.83±0.031^a,b,e
CM-25%	71.45±1.26^a,b,d	0.71±0.043^a,b,d
CM-50%	66.34±1.40^a,b	0.75±0.041^a,b

Groups	MII (n)	IVF (%)	Cleavage (%)	Morula (%)	Blastocyst (%)	Degenerated (%)
Normal
Co	120	72.66±1.13^b,c,d,e,f	60.46±1.14^b,c,d,e,f	7.72±1.01	47.82±0.66^b,c,d,e,f	5.21±1.09
RG	124	78.74±1.10^a,d,e,f	65.24±0.83^a,d,e,f	4.31±0.95	54.48±0.52^a,d,e,f	5.36±0.35
LC-0.3	122	79.16±0.96^a,e,f	68.84±0.93^a,e,f	4.16±1.17	59.11±1.04^a,d,e,f	3.25±0.61
LC-0.6	126	85.33±1.07^a,b,f	72.38±1.05^a,b,f	5.28±0.91	63.16±0.74^a,b,c,f	3.84±0.95
CM-25%	124	88.90±0.88^a,b,c	77.56±0.75^a,b,c,f	6.14±0.48	67.55±0.87^a,b,c	3.78±0.74
CM-50%	122	93.77±0.92^a,b,c,d	84.81±0.44^a,b,c,d,e	7.20±0.62	73.19±0.82^a,b,c,d	4.32±1.04
Endometriosis
Co	134	70.00±0.25^d,e,f	50.13±0.08^b,c,d,e,f	10.23±0.03^b,c,d,e,f	18.09±0.10^c,d,e,f	13.14±0.17^d,e,f
RG	130	71.32±0.04^d,e,f	57.17±0.05^a,c,d,e,f	16.14±0.17^a,d,e,f	19.27±0.34^c,d,e,f	11.06±0.19
LC-0.3	128	72.58±0.10^d,e,f	63.54±0.20^a,b,e,f	18.29±0.38^a,d	26.33±0.11^a,b,d,e,f	9.21±0.05
LC-0.6	131	81.20±0.05^a,b,c,f	65.45±0.34^a,b,e,f	23.19±0.18^a,b	34.40±0.32^a,b,c,e,f	8.05±0.14^a
CM-25%	136	83.31±0.17^a,b,c,f	73.25±0.11^a,b,c,d	21.06±0.20^a,b	45.05±0.19^a,b,c,d,f	8.00±0.16^a
CM-50%	132	91.59±0.0^a,b,c,d,e	78.63±0.14^a,b,c,d	24.59±0.41^a,b,c	53.28±0.24^a,b,c,d,e	8.04±0.11^a

Groups	Blastocysts (n)	Total cells (n)	TE (n)	ICM (n)
Normal
Co	25	51.24±0.35^b,c,d,e,f	37.82±0.61^c,d,e,f	13.26±0.28
RG	25	56.33±0.44^a,e,f	43.16±0.52^f	13.04±0.61
LC-0.3	25	57.82±0.58^a,e,f	44.77±0.94^a	13.01±0.35
LC-0.6	25	59.36±0.48^a,f	44.98±0.28^a	14.30±0.73
CM-25%	25	62.95±0.88^a,b,c	46.52±0.66^a	16.41±0.80
CM-50%	25	65.87±0.56^a,b,c,d	47.24±0.76^a,b	18.57±0.25^a
Endometriosis
Co	25	47.12±0.91^c,d,e,f	36.11±1.07^c,d,e,f	10.90±0.39
RG	25	48.02±0.67^d,e,f	36.24±0.78^c,d,e,f	11.18±0.56
LC-0.3	25	52.75±1.04^a,f	41.35±0.56^a,b	11.33±0.42
LC-0.6	25	54.36±0.48^a,b	41.94±0.28^a,b	12.35±1.02
CM-25%	25	54.84±0.72^a,b	43.78±0.44^a,b	11.06±0.47
CM-50%	25	57.25±0.81^a,b,c	44.89±0.93^a,b	12.34±0.66

Associação Brasileira de Divulgação Científica Av. Bandeirantes, 3900, 14049-900 Ribeirão Preto SP Brazil, Tel. / Fax: +55 16 3315-9120 - Ribeirão Preto - SP - Brazil
E-mail: bjournal@terra.com.br

Acompanhe os números deste periódico no seu leitor de RSS

[1] Correspondence: Mojtaba Moradi: <mojimovt@gmail.com> | Mehri Azadbakht: <azadbakhtm_tmu@yahoo.com>